Free Essay

Research

In: Science

Submitted By Pooja1404
Words 4190
Pages 17
International Journal of Biotechnology. Photon 114 (2015) 444-451 https://sites.google.com/site/photonfoundationorganization/home/international-journal-of-biotechnology Original Research Article. ISJN: 3352-7304: Impact Index: 4.23

International Journal of Biotechnology

Ph ton

Study of Microbial Beta-Galactosidase Isolated from Fermented Millets
Mixture
Pooja Jagtap, Humera Bhattiwala and Annika Durve*
Department of Biotechnology, Birla College of Arts, Science and Commerce, Kalyan. Dist Thane, 421301, India
Article history:
Received: 31 March, 2015
Accepted: 07 April, 2015
Available online: 28 September, 2015
Keywords:
Beta: galactosidase, ONPG, X: gal, ONPG and X: gal
Corresponding Author:
Durve A.*
Associate Professor
Email: annikadurve ( at ) yahoo ( dot ) com
Bhattiwala H.
Lecturer
Jagtap P.
Scientific Officer

Abstract
In the present study microorganisms producing betagalactosidase enzyme were isolated from fermented millets mixture. Three bacterial isolate C2, C4, Y were found to be Gram positive bacilli, Gram negative cocobacili, Gram positive bacilli respectively while isolate SC2 was found to be yeast. These bacterial

isolates were identified using various biochemical tests
Growth study kinetics of microbial isolates was performed and the optimum pH and temperature was found. Beta-galactosidase enzyme was extracted and the effect of pH, temperature, substrate variation and incubation period on enzyme activity was studied. The activity of beta-galactosidase for all the microbial isolate increased with increase in temperature upto 37°C.
Michaelis-Menten plot and Lineweaver Burk plot were constructed to calculate the Km and Vmax values.
Immobilization of crude enzyme from the isolates was performed and enzyme activity using ONPG and lactose as a substrate was found. SDS PAGE analysis of the dialysate sample of bacterial isolate C2 and yeast isolate
SC2 was carried out. A band having molecular weight of approximately 70-80 kDa was detected.
Citation:
Jagtap P., Bhattiwala H., Durve A., 2015. Study of Microbial
Beta-Galactosidase Isolated from Fermented Millets Mixture.
Photon 114, 444-451
All Rights Reserved with Photon.
Photon Ignitor: ISJN33527304D785828092015

1. Introduction
Lactose is a diasacharide sugar derived from glucose and galactose which is found in milk.
Lactose makes up around 2-8% of milk (by weight) although the amount varies among species and individuals. Lactose intolerance primarily refers to a syndrome having one or more symptoms upon the consumption of food substances containing lactose.
Individuals may be lactose intolerant to varying degrees, depending on the severity of these symptoms. Lactose malabsorption refers to the physiological concomitant of lactase deficiency
(i.e., the body does not have sufficient lactase capacity to digest the amount of lactose ingested).
A medical condition with similar symptoms is fructose malabsorption.
Lactose intolerance is not an allergy because it is not an immune response, but rather a problem with
Ph ton

digestion caused by lactase deficiency. Milk allergy is a separate condition, with distinct symptoms that occur when the presence of milk proteins trigger an immune reaction.
Lactose intolerance is usually caused by a deficiency of the enzyme called lactase (a protein that causes a chemical reaction to occur). A lactase substitute is available that can be taken to replace the lactase that your body cannot produce. The lactase substitute comes in liquid form (usually as drops) that can be taken before a meal or added to milk. Lactase enzyme, in the form of drops and capsules, are available from most health foods shops (Food Standards Agency, health-library articles, 2011).
The enzyme Beta Galactosidase (β-gal, EC
3.2.1.23) belongs to the group of hydrolases, forming two products from one substrate. The

444

enzyme catalyzes the hydrolysis and transgalactosylation reaction of β-D-galactopyranosides.
These enzymes are found widespread in nature: microorganisms, plants, and animals. Microbial
Beta Galactosidases are of major interest due to their easy application in bioprocess technology.
The microbial enzymes have long been used for the hydrolysis of lactose, for increasing the digestibility of milk, or for improving the functional properties of dairy products. During the past decades, the Beta Galactosidase-catalyzed transgalactosylation has proved to be useful for structural and functional modification of food materials, medicines, and other biologically active compounds. Additionally, microbial Beta
Galactosidase are taken as a marker enzyme for coliform bacteria which are indicators of the faecal pollution of water (Nakayama and Amachi, 1999).
In nature, the substrate for Beta Galactosidase is lactose. Many chromogenic, fluorogenic, and luminogenic substrates that are specific for Beta
Galactosidase have been developed both substrate structures together with their respective reaction products are displayed (Nakayama and Amachi,
1999).
The aim of the present work was to isolate microbial sources capable of hydrolyzing a wide range of lactose. Bacteria and yeasts were isolated from fermented mixture of millets. The ability of these microorganisms to grow and produce lactase
(Beta Galactosidase) under different environmental conditions was studied. The microorganisms were capable of producing lactase in the presence of lactose. Various environmental conditions viz. effect of pH, temperature, enzyme concentration and substrate concentration were evaluated and the optimum conditions for the production of lactase were studied.
2. Material and Methods
Millets which included jowar (Sorghum), ragi
(Eleuisine coracana), bajra (Pennisetum glaucum) was purchased from local market. All the chemicals, reagents and media were prepared in distilled water. The prepared media was autoclaved at 15 lbs at 121.7˚C for 20 minutes.
2.1 Isolation, screening and identification of microorganisms producing β-Galactosidase
The millets were soaked in water and incubated at room temperature for 3 days. The millets were crushed with a mortar and pestle. The supernatant was streaked on MRS medium (De Man et al.,
1960) containing 2% lactose. The microorganisms obtained were screened for β-Galactosidase enzyme using Luriya Bertani agar plates infused with 50 µl X-Gal (5-bromo-4-chloro-3-indole-βPh ton

Dgalactopyranoside; 20mg/ml of DMSO) and 10 µl
IPTG (iso-propyl-thio-β-D-galactopyranoside) as an inducer to select the colonies showing lactose fermenting ability. The plates were then incubated at 37ºC for 24 to 48 hrs. The microrganisms were identified using Gram staining and biochemical characteristics (Harley and Prescott, 2002). The effect of pH, temperature and growth kinetics was studied. 2.2. Beta-galactosidase activity measurement using the ONPG standard assay
The enzyme extraction was carried out using toluene/acetone (1:9 v/v) solution and vortexed.
The supernatant was used as a crude enzyme source (Gheytanchi et al., 2010).
The activity assay was carried out by adding 200µL of enzyme extract and 900µL 50mM phosphate buffer (pH 6.5) along with 300µl of 0.132mM
ONPG solution. The reaction mixture was incubated for 15mins at 37°C. The reaction was stopped by adding 1000 µL 0.4 M Na2CO3 solution. The absorption of the yellow colour was measured photo metrically at 420 nm.
The calculation of the enzyme activity was carried out as described by (Nguyen et al., 2006). One unit
(U) of ONPG activity is defined as the amount of enzyme releasing 1 µ mol of ONP per minute under the specific assay conditions.
2.3 Determination of the ONP standard curve
Concentrations of ONP were prepared in sodium phosphate buffer, and the absorption was read at
420 nm, as described by Nguyen (2006). The standard curve was determined.
2.4 Lactose and glucose assay
Beta-galactosidase activity was determined by using lactose as substrate. 400µl of enzyme extract was added to 600µl of 10% and 15% lactose, separately and incubated for 15 minutes. When the mixture was cooled to room temperature, the release of D-glucose was determined photo metrically by using the standard GOD/POD assay
(Nguyen et al., 2006). One unit (U) of betagalactosidase activity is defined as the amount of enzyme releasing 1 µmol of D-glucose per minute under the described conditions.
Glucose concentrations ranging from 0.1- 1g/L were prepared in water and glucose concentration was determined by GOD-POD assay (ERBA GODPOD assay kit). The standard curve was determined. 445

2.5. Protein content
Protein content was found by Folin Lowry method
(Lowry, 1951) using bovine serum albumin as
Standard in range of (100µg-1000µg/ml).
2.6. Enzyme kinetics
The enzyme kinetics was studied for the crude enzymes by determining the optimum pH, temperature, enzyme concentration and substrate concentration. 2.6.1 Determination of pH optimum
To determine the optimum pH for the enzyme, the reaction mixture of enzyme and substrate was incubated in set of buffer of a pH ranging from 4 to
8.5 at 37ºC for 30 minutes. The activity of the betagalactosidase at different pH was determined. The
ONPG standard assay was performed using different pH (Nath et al., 2013; Natrajan et al.,
2012).
2.6.2 Determination of optimum temperature
The optimum temperature for enzyme activity was determined by varying the incubation temperature of the reaction mixture at 4ºC, 28ºC, 37ºC and 55ºC.
The activity of the beta-galactosidase at different temperatures was determined. The ONPG standard assay was performed using different incubation temperatures (Nath et al., 2013; Natrajan et al.,
2012).

2.9 Immobilization
The two isolates which showed maximum enzyme activity using ONPG substrate were immobilized in
Sodium alginate (Ayaz et al., 2011). This involved mixing of extracted enzyme from 100ml of overnight grown culture of the two isolates with
3% alginate. With the help of a syringe this was then dropped drop wise into 6% chilled CaCl2 that resulted in the formation of beads. The beads so formed were cured overnight at 4˚C. The beads were activated by incubating in sterile MRS medium containing 2 % lactose overnight. These beads were then introduced into 10ml of 4mg/ml
ONPG and standard protocol was used to find enzyme activity (Ayaz et al., 2011). Also beads were introduced in 10ml of 10% and 15% lactose, separately and release glucose was found by GODPOD assay (ERBA GOD-POD assay kit).
2.10 Application of Beta Galactosidase- Use of
Lactase in Bread Making
To 80g of flour, 332ml of yeast suspension (7.5g of baker’s yeast suspended in 95 ml of water) and 48 ml of a lactase (beta-galactosidase) enzyme 5ml of milk and sugar was added. The contents were mixed thoroughly with a spatula. After mixing to optimum consistency, the doughs were fermented at 30°C (Husain, 2010).
3. Results and Discussion

2.6.3 Determination of optimum substrate concentration To determine optimum substrate concentration, various concentration of substrate (ONPG) ranging from 0.016, 0.033, 0.049, 0.066, 0.082, 0.099,
0.132 mM, were used and the reactions were carried out at 37ºC for 30minutes. The activity of the beta-galactosidase at different concentration of substrate was determined. The ONPG standard assay was performed using different substrate concentration (Nath et al., 2013; Natrajan et al.,
2012).

3.1 Isolation, screening and identification of microorganisms producing β-Galactosidase
Beta-galctosidase producing colonies were selected by blue white selection method by growing them in the presence of IPTG and X-Gal
(Fig 3.1). Four different beta-galactosidase producing colonies were isolated. The four different colonies which were X-Gal positive were tested for utilization of ONPG substrate. All four colonies were able to break ONPG into ONP, which is a yellow colour compound. Gram staining and colony characters were studied.

2.7. Effect of incubation time on microbial cells to obtain crude enzyme
The effect of incubation period, on betagalactosidase production was investigated by cultivating the organism at different incubation time (24-96 hours). The organism was incubated, the beta-galactosidase activity was determined in supernatant. Figure 3.1: Blue colonies producing Beta-galctosidase

indicating

organisms

2.8 SDS-PAGE analysis
Beta galctosidase enzyme was partially purified by
Ammonium sulphate precipitation followed by dialysis. The enzyme was characterised using SDSPAGE and the molecular weight was determined.
Three bacterial isolate C2, C4, Y were found to be
Gram positive bacilli, Gram negative cocobacili,
Ph ton

446

Gram positive bacilli respectively while isolate
SC2 was found to be yeast. The bacterial isolates were subjected to various biochemical tests and were identified according to Bergey’s manual of
Systematic Bacteriology (Volume I and II).
Standard Biochemical were Indole, Methyl Red,
Vogues Proskauer and Citrate Test (IMViC);
Triple Sugar Iron slant test and sugar fermentation containing 1% sugar solutions of sucrose, glucose, lactose, xylose, maltose, mannitol, mannose and fructose containing Andrade’s indicator with an inverted Durham’s tubes. These isolates were identified till species level using biochemical tests.
On the basis of the biochemical results, these isolates were found to belong to Bacilli sp.,
Lactobacillisp. and Vibrio sp.
The optimum pH for growth for bacterial isolate was found to be 7 while for yeast was pH 5. The optimum temperature for all microbial isolates was found to be 37°C. The microbial isolates could grow even in the presence of NaCl. The isolates could tolerate NaCl concentration upto 6.5% for isolate C2, 3 % for isolate C4 and Y and 12.5 % for
SC2.

enzyme activity was found to be 0.0266 enzyme activity /ml. At pH 7.5 for yeast isolate SC2 enzyme activity was found to be 0.0366 enzyme activity /ml (Table 3.1; Fig 3.2).
3.2.2 Determination of Temperature
The optimum temperature required for betagalactosidase activity was checked by incubating the assay reaction at different temperature viz. 4°C,
28°C, 37°C and 55°C. The activity of betagalactosidase for all the microbial bacterial isolate increased with increase in temperature upto 37°C
(Fig 3.3). However, further rise in reaction temperature caused loss of beta-galactosidase activity (Table 3.2).
Temp
(°C)
4
28
37
55

Enzyme activity/ml
C2
C4
0.0033
0.0011
0.0122
0.0044
0.03
0.0111
0.0111
0.0022

Y
0.0033
0.0111
0.0166
0.0044

SC2
0.0077
0.02
0.0355
0.01

Figure 3.3: Effect of temperature on Enzyme activity

3.2 Enzyme kinetics
Beta galactosidase was extracted by treating the culture broth with toluene: acetone (1:9) and the supernatant collected was used as crude enzyme.
The various parameters i.e. effect of pH, temperature, enzyme concentration and substrate concentrations for the crude enzyme were determined. 3.2.1 Determination of pH
The enzyme activity for C2 was found to be 0.029 enzyme activity/ml at pH 5; for C4 bacterial isolate, enzyme activity was found to be
0.0111enzyme activity/ml respectively. The elevation of pH beyond this caused rapid decrease in enzyme activity. At pH 7, for bacterial isolate Y pH 4
5
5.5
6
6.5
7
7.5
8
8.5
9

Enzyme activity/ml
C2
C4
0.027
0.0011
0.029
0.0022
0.0024
0.0066
0.02
0.0077
0.014
0.0111
0.016
0.0099
0.01
0.0088
0.0088
0.0055
0.0033
0.0022
0.0011
0.0022

Ph ton

Y
0.0044
0.0044
0.0077
0.0111
0.0177
0.0266
0.0088
0.0033
0.0011
0.0011

SC2
0.015
0.019
0.033
0.026
0.019
0.010
0.007
0.005
0.004
0.001

3.2.3 Determination of Substrate variation
The effect of substrate concentration was checked on beta-galactosidase activity. Kinetic constants i.e.
Km and Vmax of the beta-galactosidase enzyme were determined by changing the ONPG substrate concentration. ONPG substrate concentrations in the assay medium were ranging from 0.0016mM to
0.0132mM. Michaelis-Menten plot and Lineweaver
Burk plot were constructed to calculate the Km and
Vmax values. Km and Vmax values for the four microbial isolate were found to be 0.0231, 0.0808,
0.0366, 0.0366, 0.0319 mM and 0.0244, 0.1009,
0.0368, 0.0328 U/mg protein respectively (Fig 3.4).
Figure 3.4: Effect of substrate variation on beta galactosidase enzyme using ONPG as substrate
(Michaelis-Menten graph).

447

3.2.4 Effect of incubation time on microbial cells to obtain crude enzyme
According to the results taken at regular intervals, the maximum enzyme activity was obtained at 48th hour of incubation. Beyond this, the enzyme productivity was remains decreased and no further increase in production was observed (Fig. 3.5). It might be due to decrease in the nutrient availabitity in the medium, or catalobic repression of the enzyme. Figure 3.5: Effect of Incubation time on microbial cells to obtain crude enzyme

3.3.1 Lactose assay
Bacterial isolate C2 and yeast isolate SC2 showed highest enzyme activity using ONPG as a substrate so, this two isolate were used to find enzyme activity using lactose as a substrate. Lactose hydrolysis was done using 10% and 15% lactose and released glucose was estimated using god-pod assay (ERBA-God Pod assay kit).
The bacterial isolate C2 which showed highest enzyme activity using ONPG as substrate was used to find enzyme activity of beta-galactosidase using lactose as a substrate (10% and 15%) and comparative study was made using yeast isolate
SC2. Bacterial isolate C2 showed enzyme activity of 0.404µmole/minute and 0.809µmole/minute using 10% and 15% lactose respectively. Similarly, yeast isolate SC2 showed enzyme activity of
2.83µmole/minute and 3.23µmole/minute using
10% and 15% lactose respectively (Table 3.3).
Yeast isolate SC2 showed higher enzyme activity than bacterial isolate at both 10% and 15% lactose as substrate (Fig: 3.6 and Fig 3.7).

The optimum pH for the Lactobacillus acidophilus enzyme was found to be 6.5-7.0 (Akolkar et al.,
2005), pH 6.5 for Thermophile B1.2 (Osiriphun and Jaturapiree, 2010) and pH 7 for Bacillus
Sp.MPTK121 (Mukeshkumar et al., 2012).
The optimum temperature for the Lactobacillus acidophilus enzyme was found to be 50°C (Akolkar et al., 2005), 60°C for Thermophile B1.2
(Osiriphun and Jaturapiree, 2010) and 30°C
Bacillus Sp.MPTK121 (Mukeshkumar et al., 2012).

Table 3.3: Enzyme activity using lactose as a substrate
Isolate
Lactose %

10
15

Enzyme activity (µmole/minute)
C2
SC2
0.404
2.83
0.809
3.23

Figure 3.6: Lactose assay for the isolate C2 and SC2 isolate using 10% lactose

Using ONPG as a substrate Km value for the
Lactobacillus acidophilus enzyme was found to be
0.11mM (Akolkar et al., 2005), 28.85mM for
Thermophile B1.2 (Osiriphun and Jaturapiree,
2010) while Vmax value for the Lactobacillus acidophilus enzyme was found to be 4.94U/mg protein (Akolkar et al., 2005) and 8.38 U/mg protein for Thermophile B1.2 (Osiriphun and
Jaturapiree, 2010).
The maximum enzyme production of beta galactosidase from Bacillus Sp.MPTK 121 was obtained at 48th hour of incubation. Beyond this, the enzyme productivity remained constant and no further increase enzyme production was observed
(Mukeshkumar et al., 2012).

Figure 3.7: Lactose assay for the isolate C2 and SC2 isolate using 15% lactose

3.3 Partial Purification of beta-galactosidase
Enzyme precipitation was carried out by salting out with ammonium sulphate. Further partial purification of beta-galactosidase was carried out by the process of dialysis using 10kDa dialysis bag.

Ph ton

448

3.4 Immobilization
Immobilized enzyme was used to find enzyme activity using ONPG (4mg/ml) and lactose (10% and 15%) as substrate. A comparative study was made of free and immobilized enzyme using lactose as substrate over time duration of 1 hour with 15minutes interval (Fig. 3.8).
Figure 3.8: (A) Immobilized beads having crude betagalactosidase from C2
(B) Immobilized beads having crude beta-galactosidase from SC2

A

yeast and bacterial isolate SC2 and C2 in Free State showed increased dissociated glucose as time increased, while in immobilized state the same enzyme showed increased dissociated glucose as time increased (Fig 3.10 and Fig 3.11). It was found that the immobilized enzyme from bacteria and yeast was more effective than compared to its free form.
Figure 3.10: Comparative study of immobilized and free crude beta-galactosidase from isolate C2 using 10% and
15% lactose

B

3.4.1 ONPG assay
Using ONPG as a substrate, immobilized crude beta-galactosidase from both C2 and SC2 isolate was used to find amount of dissociated ONP which is equal to optical density of reaction mixture at
420nm. O.D of reaction mixture was found at interval of 15 minutes till 60 minutes. At 60 minute both immobilized enzyme from C2 and SC2 isolate showed highest O.D (Fig 3.9). The immobilized yeast enzyme SC2 showed higher dissociated ONP than bacterial isolate C2 enzyme.

Figure 3.11: Immobilized and free crude betagalactosidase from isolate SC2 using 10% and 15% lactose Figure 3.9: ONPG assay using immobilised crude betagalactosidase from both C2 and SC2 isolate

3.4.2 Lactose assay
Using lactose (10% and 15%) as a substrate immobilized crude beta-galactosidase from both C2 and SC2 isolate was used to find amount of dissociated glucose which is equal to optical density of reaction mixture at 505nm. Comparative study of free and immobilized enzyme was done. It was found that crude beta-galactosidase from both
Ph ton

3.5 Electrophoresis and Molecular Weight
Determination by SDS-PAGE
SDS Page analysis of the purified betagalactosidase from C2 and SC2 isolate was carried out. A molecular ladder was run along with the sample. The PAGE analysis showed a band having molecular weight of approximately70-90kDa for bacteria n yeast (Fig 3.12). References have show that the molecular weight of beta-galactosidase enzyme have a molecular weight of 70-75kDa
(Mukeshkumar et al., 2012; Osiriphun and
Jataurapiree, 2010).
3.6 Application- Use of beta-galactosidase in bread making The enzyme from both bacterial isolate C2 and yeast isolate SC2 was use in bread making. The dough containing enzyme and control dough

449

having no enzyme was fermented and after 40 mins, the dough containing the enzyme was found to be softer than the control and the texture of the bread was found to be softer compared to the
Figure 3.12: SDS PAGE analysis of crude betagalactosidase enzyme from C2 bacterial isolate and
SC2 yeast isolate

to produce beta-galactosidase enzyme. The enzyme obtained from the bacteria was found to be more effective than the enzyme obtained from yeast. The bacterial isolates were able to grow in acidic environment and showed high enzyme activity at acidic pH which can be used at industrial scale.
These isolates on further strain improvement can be used for producing beta galactosidase commercially.
References
Akolkar S., Sajgure A., Leele S. 2006. β-galactosidase from lactobascillus acidophilus isolated from fermented ragi (Eluisene coracana).Indian journal of biotechnology.5, 184-188.
Ayaz N., Alnahdi H., Danial E. 2011. Immobilization and Properties of β-D-Galactosidase from Bacillus
Licheniformis E66. Journal of Applied Sciences
Research.7(12), 2448-2454

control. The enzyme obtained from the bacteria was found to be more effective than the enzyme obtained Figure 3.13: Dough containing enzyme (C2 and SC2) and no enzyme (control)

Ayaz N., Alnahdi H., Danial E. 2011. Immobilization and Properties of β-D-Galactosidase from Bacillus
Licheniformis E66. Journal of Applied Sciences
Research. 7(12), 2448-2454.
De Man J.C., Rogosa M., Sharpe M.E. 1960. A medium for the cultivation of lactobacilli. J. Applied Bacteriol.
23, 130-135.
Food Standards Agency,health-library articles., 2011.
Gheytanchi E., Heshmati F., Shargh B.E., Nowrojee J.,
Movahedzadeh F. 2010. Study on beta galactosidase enzyme produced by isolated lactobacilli from milk and cheese. Applied journal of microbiology Research. 4(6),
454-458.
Harley J.P., Prescott L. M. 2002. Laboratory exercises in microbiology. Fifth edition. The McGraw –Hill publishing companies.

Figure 3.14: Texture of bread

Husain Q. 2010. β Galactosidases and their potential applications: a review. Critical Reviews in
Biotechnology. 30(1), 41–62.
Kumar M., Sudha M., Devika S., Balakumaran M.,
Kumar R., Kalaichelvan P. 2013. Production and
Optimization of β-galactosidase by Bacillus Sp. MPTK
121, Isolated from Dairy Plant Soil. Annals of Biological
Research. 3(4), 1712-1718.
Lowry O. H., Rosebrough J., Farr A. L., Randall R. J.
1951. Protein measurement with the folin phenol reagent.
Journal of Biological Chemistry. 193, 265–275.

Conclusion
In the present study microorganisms producing beta-galactosidase enzyme were isolated from fermented millets mixture. Three bacterial isolates and yeast were selected on the basis of their ability
Ph ton

Nakayama T., Amachi T. 1999. β-Galactosidase, enzymology. In:
Encyclopedia
of Bioprocess
Technology:
Fermentation,
Biocatalysis,
and
Bioseparation (Flickinger MC, Drew SW, editors), John
Wiley and Sons, New York, NY .3, pp, 1291-1305.
Nath A., Sarkar S., Maitra M., Bhattacharjee C.,
Chowdhury R. 2013. Production, Purification and
Characterization of β-galactosidase Synthesized by

450

Bacillus safensis (JUCHE1). Advances in Biotechnology and Pharmaceutical Sciences.7-16.
Natrajan J., Christobell C., Kumar M., Kumar R.,
Balakumaran M., Kalaichelvan P. 2012. Isolation and
Characterization of β-D-Galactosidase Producing
Bacillus sp. from Dairy Effluent. World Applied
Sciences Journal.17 (11): 1466-1474.
Nguyen T.H., Splechtna B., Haltrich D. Kulbe K. D.,
Peterbauer C. 2007. Cloning and expression of beta galacosidase genes from lactobacillus reuteri in
Escherichia coli. Journal of Biotechnology. 282, 10831091.
Osiriphun S., Jaturapiree P. 2009. Isolation and characterization of β-galactosidase from thermophile
B1.2.Asian journal of food and agro-industry. 2(4), 135143.

For publications/ Enquiries/ Submissions:
Email: photonjournal@yahoo.com
Ph ton

451

Similar Documents

Premium Essay

Research

...The Research Process Writers usually treat the research task as a sequential process involving several clearly defined steps. No one claims that research requires completion of each step before going to the next. Recycling, circumventing, and skipping occur. Some steps are begun out of sequence, some are carried out simultaneously, and some may be omitted. Despite these variations, the idea of a sequence is useful for developing a project and for keeping the project orderly as it unfolds. Exhibit 3–1 models the sequence of the research process. We refer to it often as we discuss each step in subsequent chapters. Our discussion of the questions that guide project planning and data gathering is incorporated into the model (see the elements within the pyramid in Exhibit 3–1 and compare them with Exhibit 3–2). Exhibit 3–1 also organizes this chapter and introduces the remainder of the book. The research process begins much as the vignette suggests. A management dilemma triggers the need for a decision. For MindWriter, a growing number of complaints about postpurchase service started the process. In other situations, a controversy arises, a major commitment of resources is called for, or conditions in the environment signal the need for a decision. For MindWriter, the critical event could have been the introduction by a competitor of new technology that would revolutionize the processing speed of laptops. Such events cause managers to reconsider their purposes or...

Words: 376 - Pages: 2

Premium Essay

Research

...requirements of talking on the phone.  These predictions were derived from basic theories on limited attention capacities. 2. Microsoft Company has basic research sites in Redmond, Washington, Tokoyo, Japan etc.at these research sites work on fundamental problems that underlie the design of future products. For example a group at Redmond is working natural language recognition soft wares, while another works on artificial intelligence. These research centres don’t produce new products rather they produce the technology that is used to enhance existing products or help new products. The product are produced by dedicates product groups. Customization of the products to match the needs of local markets is sometimes carried out at local subsidiaries. Thus, the Chinese subsidiary in Singapore will do basic customizations of programs such as MS office adding Chinese characters and customizing the interface. APPLIED RESEARCH * INTERNATIONAL TELECOMMUNICATION AND INFORMATION TECHNOLOGY SERVICE COMPANY Offering customers products and services for ‘connected life and work’  Project: 1. Research focused on the organisation’s tendency to appoint ‘safe’ senior executives, rather than those who were able to drive change through the business, and enable a culture of calculated business risk and growth.  This research led to a programme that created different and improved relationships with executive search agencies, established a positive shift in the interaction between the...

Words: 282 - Pages: 2

Premium Essay

Research

...Myresearch About 30 million other animals. Animal experimentation by scientists can be cruel and unjust, but at the same time it can provide long term benefits for humanity. Animals used in research and experiments have been going on for 2,000 years and keep is going strong. It is a widely debated about topic all over the world. Some say it is inhuman while others say it's for the good of human kind. There are many different reasons why people perform experiments and why others total disagree with it. Each year 20 million animals are produce and breed for the only purpose but to be tested on. Fifty-three thousands of animals are used each year in medical and veterinary schools. The rest is used in basic research. The demand for animals in the United States is 50 million mice, 20 million rats, and aThis includes 200,000 cats and 450,000 dogs. The world uses about 200-250 million animals per year. The problem with working with animals is that they cannot communicate their feelings and reactions. Other people say that they can communicate and react to humans just a well as one person to another. Some of the animals the research's use are not domesticated which makes them extremely hard to control and handle. The experiments that go on behind closed doors are some of the most horrific things a human could think of too torture somebody or something. Animals in labs are literally used as models and are poked at and cut open like nothing is happening. When drug are tested on animals...

Words: 355 - Pages: 2

Premium Essay

Research

...Research Methodology & Fundamentals of MR. 100 Marks Course Content 1. Relevance & Scope of Research in Management. 2. Steps Involved in the Research Process 3. Identification of Research Problem. 4. Defining MR problems 5. Research Design 6. Data – Collection Methodology, Primary Data – Collection Methods / Measurement Techniques – Characteristics of Measurement Techniques – Reliability, Validity etc. – Secondary Data Collection Methods – Library Research, References – Bibliography, Abstracts, etc. 7. Primary and Secondary data sources and data collection instruments including in-depth interviews, projective techniques and focus groups 8. Data management plan – Sampling & measurement 9. Data analysis – Tabulation, SPSS applications data base, testing for association 10. Analysis Techniques – Qualitative & Quantitative Analysis Techniques – Techniques of Testing Hypothesis – Chi-square, T-test, Correlation & Regression Analysis, Analysis of Variance, etc. – Making Choice of an Appropriate Analysis Technique. 11. Research Report Writing. 12. .Computer Aided Research Methodology – use of SPSS packages Reference Text 1. Business Research Methods – Cooper Schindler 2. Research Methodology Methods & Techniques – C.R.Kothari 3. D. K. Bhattacharya: Research Methodology (Excel) 4. P. C. Tripathy: A text book of Research Methodology in...

Words: 5115 - Pages: 21

Premium Essay

Research

...Research Research is a systematic inquiry to describe, explain, predict and control the observed phenomenon. Research involves inductive and deductive methods (Babbie, 1998). Inductive methods analyze the observed phenomenon and identify the general principles, structures, or processes underlying the phenomenon observed; deductive methods verify the hypothesized principles through observations. The purposes are different: one is to develop explanations, and the other is to test the validity of the explanations. One thing that we have to pay attention to research is that the heart of the research is not on statistics, but the thinking behind the research. How we really want to find out, how we build arguments about ideas and concepts, and what evidence that we can support to persuade people to accept our arguments. Gall, Borg and Gall (1996) proposed four types of knowledge that research contributed to education as follows: 1. Description: Results of research can describe natural or social phenomenon, such as its form, structure, activity, change over time, relationship to other phenomena. The descriptive function of research relies on instrumentation for measurement and observations. The descriptive research results in our understanding of what happened. It sometimes produces statistical information about aspects of education. 2. Prediction: Prediction research is intended to predict a phenomenon that will occur at time Y from information at an earlier time X. In educational...

Words: 1179 - Pages: 5

Free Essay

Research

...Contents TITLE 2 INTRODUCTION 3 BACKGROUND OF THE STUDY 3 AIM 4 OBJECTIVES 4 RESEARCH QUESTIONS 4 LITERATURE REVIEW 5 METHODOLOGY AND DATACOLLECTION 5 POPULATION AND SAMPLING 6 DATA ANALYSIS METHODS 6 PARTICIPANTS IN THE STUDY 7 STUDY PERIOD (GANTT CHART) 8 STUDY RESOURCES 9 REFERENCES 9 BIBLIOGRAPHY 9 APPENDICES: 10 * The Impact of Motivation through Incentives for a better Performance - Adaaran Select Meedhupparu Ahmed Anwar Athifa Ibrahim (Academic Supervisor) Applied Research Project to the Faculty of Hospitality and Tourism Studies The Maldives National University * * Introduction As it is clear, staff motivation is important in all the sectors especially in the tourism sector where we require highly skilled employees to get the best of their output to reach the organizational goals. Therefore, organizations spend a lot on their staff motivation in terms of different incentive approaches, such as financial benefits, training and development, appreciations, rewards and promotions. As mentioned in the title, the outline of the findings will be focused on the motivation of the staffs on improving their performances by the different incentive packages that they get at the resort. This study will be executed at Adaaran Meedhupparu by giving questionnaire to the staff working in different departments to fill up and return to the scholar to examine the current situation of staff satisfaction on motivation to do...

Words: 2768 - Pages: 12

Premium Essay

Research

...STEP 1etasblish the need for research We have to consider if it is a real need for doing a research? Research takes time and costs money. If the information is already available, decisions must be made now, we cant afford research and costs outweigh the value of the research, then the research is not needed. Step 2 define the problem or topic State your topic as a question. This is the most important step. Identify the main concepts or keywords in your question. Problem maybe either specific or general. Step 3 establish research objective Research objectives, when achieved, provide the Information necessary to solve the problem identified in Step 2. Research objectives state what the researchers must do. Crystallize the research problems and translate them into research objective. At this point, we will pin down the research question. Step 4 determine research design The research design is a plan or framework for conducting the study and collecting data. It is defined as the specific methods and procedures you use to acquire the information you need. based on the research objectives, we will determine the most appropriate research design: qualitative and/ or quantitative. • Exploratory Research: collecting information in an unstructured and informal manner. • Descriptive Research refers to a set of methods and procedures describing marketing variables. • Causal Research (experiments): allows isolation of causes and...

Words: 1099 - Pages: 5

Premium Essay

Research

...Research Article Research is important to every business because of the information it provides. There is a basic process to researching information and that process begins by deciding what information needs to be researched. The next step is to develop a hypothesis, which describes what the research paper is about and what the researcher’s opinion is regarding the topic. The research article chosen for this paper is titled, “The Anchor Contraction Effect in International Marketing Research.” The hypothesis for this paper is, “This raises the issue of whether providing responses on rating scales in a person’s native versus second language exerts a systematic influence on the responses obtained.” Simply explained, the hypothesis of this paper is to determine whether research questions should be in a person’s native language rather than expecting them to respond to questions in a language in which they might not be fluent. The hypothesis of this paper was accepted based on the research data gathered by the research team. This hypothesis was supported by nine studies using a variety of research methods. The research methods provided data that demonstrated the level of inaccuracy based on questions being asked in a language that was not the respondent’s native language. The research data provided insight into the probability of more accurate results when the respondent was asked questions in a manner that related well with their culture. There are several implications...

Words: 322 - Pages: 2

Premium Essay

Research

...ACE8001: What do we mean by Research? & Can we hope to do genuine Social Science Research (David Harvey)  What do we mean by research? What might characterise good research practice? There is no point in us trying to re-invent the wheel - other and probably more capable people than us have wrestled with this problem before us, and it makes good sense and is good practice to learn what they have discovered.  In other words - we need to explore more reliable and effective methods and systems for the pursuit of research than we have been doing so far. What is research? Dictionary Definitions of Research: * "The act of searching closely or carefully for or after a specified thing or person" * "An investigation directed to discovery of some fact by careful study of a subject" * "A course of scientific enquiry" (where scientific = "producing demonstrative knowledge") Howard and Sharp (HS) define research as:  "seeking through methodical processes to add to bodies of knowledge by the discovery or elucidation of non-trivial facts, insights and improved understanding of situations, processes and mechanisms".  [Howard, K. and Sharp, J.A. The Management of a student research project, Gower, 1983 - a useful and practical “how to do it” guide] Two other, more recent guides are: Denscombe, Martyn, 2002, Ground rules for good research: a 10 point guide for social research,  Open University Press. Robinson Library Shelf Mark: 300.72 DEN, Level 3 (several copies)...

Words: 4067 - Pages: 17

Free Essay

Research

...solve analytic models or whatever, but they often fail to demonstrate that they have thoroughly thought about their papers—in other words, when you push them about the implicit and explicit assumptions and implications of their research models, it appears that they haven’t really given these matters much thought at all.[1] Too often they fall back on saying that they are doing what they are doing because that is the way it is done in the prior literature, which is more of an excuse than a answer. (Of course, once a researcher reaches a certain age, they all feel that youngsters aren’t as good as they were in the good old days!) Therefore, in this class we shall go beyond simply studying research in managerial accounting. For many of you, this is your first introduction to accounting research and to PhD level class. Hence, in these classes we shall also learn how to solve business problems systematically and to understand what it means to have thoroughly “thought through” a paper. We begin not with academic research, but with some real world cases, because we should never forget that ours is an applied research field: accounting research is a means towards the end of understanding business and is not an end in itself, in the way pure science research is. Developing a systematic procedure for solving a real world business problem is the starting point for developing a...

Words: 2437 - Pages: 10

Free Essay

Research

...manger know about research when the job entails managing people, products, events, environments, and the like? Answer: Research simply means a search for facts – answers to questions and solutions to problems. It is a purposive investigation. It is an organized inquiry. It seeks to find explanations to unexplained phenomenon to clarify the doubtful facts and to correct the misconceived facts. Research is the organized and systematic inquiry or investigation which provides information for solving a problem or finding answers to a complex issue. Research in business: Often, organization members want to know everything about their products, services, programs, etc. Your research plans depend on what information you need to collect in order to make major decisions about a product, service, program, etc. Research provides the needed information that guides managers to make informed decisions to successfully deal with problems. The more focused you are about your resources, products, events and environments what you want to gain by your research, the more effective and efficient you can be in your research, the shorter the time it will take you and ultimately the less it will cost you. Manager’s role in research programs of a company: Managing people is only a fraction of a manager's responsibility - they have to manage the operations of the department, and often have responsibilities towards the profitability of the organization. Knowledge of research can be very helpful...

Words: 4738 - Pages: 19

Free Essay

Research

...Volume 3, number 2 What is critical appraisal? Sponsored by an educational grant from AVENTIS Pharma Alison Hill BSC FFPHM FRCP Director, and Claire Spittlehouse BSc Business Manager, Critical Appraisal Skills Programme, Institute of Health Sciences, Oxford q Critical appraisal is the process of systematically examining research evidence to assess its validity, results and relevance before using it to inform a decision. q Critical appraisal is an essential part of evidence-based clinical practice that includes the process of systematically finding, appraising and acting on evidence of effectiveness. q Critical appraisal allows us to make sense of research evidence and thus begins to close the gap between research and practice. q Randomised controlled trials can minimise bias and use the most appropriate design for studying the effectiveness of a specific intervention or treatment. q Systematic reviews are particularly useful because they usually contain an explicit statement of the objectives, materials and methods, and should be conducted according to explicit and reproducible methodology. q Randomised controlled trials and systematic reviews are not automatically of good quality and should be appraised critically. www.evidence-based-medicine.co.uk Prescribing information is on page 8 1 What is critical appraisal What is critical appraisal? Critical appraisal is one step in the process of evidence-based clinical practice. Evidencebased clinical practice...

Words: 4659 - Pages: 19

Premium Essay

Research

...ISS334: Information Systems Security (Project Description) * Each student work alone in this project (individual project). * Each student is required to complete a mid-size project, which includes proposal, implementation, and final demo or paper. * Students will be grouped into teams for the sake of presentations at the end of the semester. Marks allocated for each individual piece of the project are as follows: a) Proposal – 10% b) Project write-up – 22% c) Presentation – 8% Projects include but are not limited to: * Research Paper * You can work on original research problems. The outcome should be a paper with original technical contribution. Your grade on this will be judged on originality, soundness of the approach, and quality of presentation.  * Example Topics: * Vulnerability Analysis * Wireless Security * Intrusion Detection  * Authentication * Access Control * Authorization * DNS Security * Digital Watermarking * New Attacks * Survey Paper * You can write a paper that surveys a particular field on information security. The outcome should be a paper that summarizes the trend in the field you have chosen. Your grade will be judged on the completeness of the survey, the quality of the trend analysis, and the quality of presentation. * Example topics: * Vulnerability Analysis * Wireless...

Words: 1209 - Pages: 5

Free Essay

Research

...Marketing Department, University of Strathclyde, Glasgow G4 0RQ, United Kingdom e-mail: 1sh@ukm.my 1,3 ABSTRACT This study examined the adequacy of using undergraduate student samples in research on online consumer attitudes by comparing the attitudes of students (n = 161) towards online retailing services with the attitudes of non-students (n = 252) towards such services. A structured questionnaire administered online was used to gather data on perceptions, satisfaction, and behavioral intentions with regard to online retailing services. The t-test results showed that, in general, students' attitude towards online retailing services is similar to that of non-students. Therefore, undergraduate students may be reasonable surrogates for consumers in research on online retailing. Keywords: internet users, electronic commerce, online consumer attitudes, online retailing services, student surrogates INTRODUCTION The usage of the internet as a communication and transaction medium in consumer markets is growing rapidly (Castells, 2000; Hart, Doherty, & EllisChadwick, 2000). In line with this expansion, consumer-based electronic commerce has become an emerging research area (e.g. Demangeot & Broderick, 2006, 2007; Teo, 2006; Tih & Ennis, 2006a, 2006b). In particular, a stream of research addressing issues related to online consumer attitudes (e.g. George, 2004; Wang, Chen, Chang, & Yang, 2007) and behaviors (see Cheung, Chan, & Limayem, 2005 for a review) has emerged. Although...

Words: 5659 - Pages: 23

Premium Essay

Research

...Importance Of Research Research is tool, which is utilized by my organizations and co-operations to have a fundamental knowledge of goods, products, and also to finding out consumer behavior. It is also a systematic investigation into the study of materials and sources inn order to establish facts and reach new conclusions. Research informed the marketers of Glidden paint because it helped them start from were the audience was which was Walmart in this case scenario. A marketer finds out what you want, and creates or finds product that fits you. Research aided the marketers of Glidden paint to come to a conclusion that not only should you hypothesize, you should also carry out experiments as well. In this case we see that the help of experiments helped them realize that Walmart’s brand name of saving money was not deterring the perceived quality of the paints. Meaning that Walmart’s cheap pricing of goods played no part in the durability of the paint. Research helped the marketers of Glidden paint realize that they could revamp the Walmart paint section, which has been ignored for years. They are confident that Glidden paint will do great numbers because they have raised awareness and created a media platform that consumers can interact with the most. Not only has research helped in satisfying consumer wants, it also gives the marketer an in-depth knowledge on the frequent changes of consumer taste. Research helped Glidden paint marketers realize...

Words: 319 - Pages: 2