Source Of Β-galactosidase Research Articles

A Review on the Various Sources of β-Galactosidase and Its Lactose Hydrolysis Property.

β-Galactosidase is a glycoside hydrolase enzyme that possesses both hydrolytic and transgalactosylation properties and has several benefits and advantages in the food and dairy industries. The catalytic process of β-galactosidase involves the transfer of a sugar residue from a glycosyl donor to an acceptor via a double-displacement mechanism. Hydrolysis prevails when water acts as an acceptor, resulting in the production of lactose-free products. Transgalactosylation prevails when lactose acts as an acceptor, resulting in the production of prebiotic oligosaccharides. β-Galactosidase is also obtained from many sources including bacteria, yeast, fungi, plants, and animals. However, depending on the origin of the β-galactosidase, the monomer composition and their bonds may differ, thereby influencing their properties and prebiotic efficacy. Thus, the increasing demand for prebiotics in the food industry and the search for new oligosaccharides have compelled researchers to search for novel sources of β-galactosidase with diverse properties. In this review, we discuss the properties, catalytic mechanisms, various sources and lactose hydrolysis properties of β-galactosidase.

Safety evaluation of the use of the non-genetically modified Hamamotoa singularis strain YIT 10047 as a source of β-galactosidase.

This assessment addresses the enzyme β-galactosidase which is not separated from the yeast cells used for its production. The β-galactosidase (β-D-galactoside galactohydrolase, EC 3.2.1.23) is produced with the non-genetically modified Hamamotoa singularis (formerly Sporobolomyces singularis) strain YIT 10047 by Yakult Pharmaceutical Industry Co., Ltd. The yeast cell suspension contains both live and dead yeast cells. It is intended to be used in the production of galacto-oligosaccharides (GOS). The final GOS products are free of viable cells of the H. singularis. Dietary exposure to the food enzyme total organic solids (TOS) was estimated to be up to 0.683mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests of the cell suspension did not indicate a safety concern. The systemic toxicity was assessed by means of a repeated dose 90-day oral toxicity study in rats. The Panelidentified a no observed adverse effect level of 296.25mg TOS/kg bw per day, the highest dose tested. This results in a margin of exposure above 434. A search for the similarity of the amino acid sequence of the β-galactosidase to known allergens was made and no matches were found. The Panelconsidered that, under the intended conditions of use, the risk of allergic reactions by dietary exposure cannot be excluded, but the likelihood for this to occur is low. Based on the data provided, the Panelconcluded that this yeast suspension used as a source of β-galactosidase does not give rise to safety concerns under the intended conditions of use.

Characterisation of a potential probiotic strain Paracoccus marcusii KGP and its application in whey bioremediation.

Whey, the main by-product obtained from the manufacture of cheese, which contains a very high organic load (mainly due to the lactose content), is not easily degradable and creates concern over environmental issues. Hydrolysis of lactose present in whey and conversion of whey lactose into valuable products such as bioethanol, sweet syrup, and animal feed offers the possibility of whey bioremediation. The increasing need for bioremediation in the dairy industry has compelled researchers to search for a novel source of β-galactosidase with diverse properties. In the present study, the bacterium Paracoccus marcusii KGP producing β-galactosidase was subjected to morphological, biochemical, and probiotic characterisation. The bacterial isolate was found to be non-pathogenic and resistant to low pH (3 and 4), bile salts (0.2%), salt (10%), pepsin (at pH 3), and pancreatin (at pH 8). Further characterisation revealed that the bacteria have a good auto-aggregation ability (40% at 24h), higher hydrophobicity (chloroform-60%, xylene-50%, and ethyl acetate-40%) and a broad spectrum of antibiotic susceptibility. The highest growth of P. marcusii KGP was achieved at pH 7 and 28°C, and the yeast extract, galactose, and MgSO4 were the best for the growth of the bacterial cells. The bacterium KGP was able to utilise whey as a substrate for its growth with good β-galactosidase production potential. Furthermore, the β-galactosidase extracted from the isolate KGP could hydrolyse 47% whey lactose efficiently at 50°C. The study thus reveals the potential application of β-galactosidase from P. marcusii KGP in whey bioremediation.

Effect of glucose depletion during the synthesis of galactooligosaccharides using a trienzymatic system

In this study the effect of glucose depletion using glucose oxidase and catalase, simultaneously to the synthesis of prebiotic galactooligosaccharides (GOS) by β-galactosidase was studied. Considering total GOS yield, a strong dependency on the source of β-galactosidase was found. Using an Aspergillus oryzae lactase, a small increase in GOS yield (from 50.0 ± 1.3 g/L to 54.1 ± 1.9 g/L) was noted. Due to the decreased rate of GOS disaccharide formation by the Kluyveromyces lactis enzyme, the total GOS yield was diminished (from 47.4 ± 0.1 g/L to 30.5 ± 1.7 g/L). However, for both enzymes, the synthesis of GOS tri- and higher oligosaccharides increased. Additionally, the total sugar content, and thus caloric value, of the final product was reduced, also resulting in a more purified GOS mixture.

Sources of β-galactosidase and its applications in food industry.

The enzyme β-galactosidases have been isolated from various sources such as bacteria, fungi, yeast, vegetables, and recombinant sources. This enzyme holds importance due to its wide applications in food industries to manufacture lactose-hydrolyzed products for lactose-intolerant people and the formation of glycosylated products. Absorption of undigested lactose in small intestine requires the activity of this enzyme; hence, the deficiency of this enzyme leads to lactose intolerance. Lactose intolerance affects around 70% of world's adult population, while the prevalence rate of lactose intolerance is 60% in Pakistan. β-Galactosidases are not only used to manufacture lactose-free products but also employed to treat whey, and used in prebiotics. This review focuses on various sources of β-galactosidase and highlights the importance of β-galactosidases in food industries.

A novel process for the production of high-purity galactooligosaccharides (GOS) using consortium of microbes

ABSTRACTGalactooligosaccharides (GOS) are nondigestible dietary fibers which have a beneficial effect on human health by promoting the growth of probiotic bacteria in the gut. In addition, other health benefits have been reported from oligosaccharides consumption such as stimulation of intestinal mobility, colon cancer prevention, mineral absorption as well as protection against certain pathogenic bacterial infections. The goal of this research was to develop an efficient biotransformation system using a consortium of microbes for the production of ≥85% pure GOS and reusing the cell biomass in repeated cycles of biotransformation. Production of GOS by lactose transgalactosylation using whole cells of Sporobolomyces singularis MTCC 5491 as a source of β-galactosidase and monosaccharides utilization by yeast isolate (NUTIDY007) were studied. For increasing the purity of GOS, growth and bioconversion parameters on the transgalactosylation by the whole cells were investigated. Further, continuous production of GOS was studied in a reactor with microfiltration membrane system. A maximum GOS purity of 42% was achieved using single culture of S. singularis. Under optimized conditions, single culture of S. singularis produced a maximum of 56% pure GOS. Addition of second culture to the reaction mixture for utilization of glucose significantly increased the GOS purity from 56% to ≥85%. The product consisted of tri- to penta-galactooligosaccharides. Trisaccharides were the main component of the reaction mixture. A maximum productivity of 10.9 g/L/hr was obtained under the optimum conditions.

METHOD JUSTIFICATION OF Β-GALACTOSIDASE ACTIVITY OF ENZYME PREPARATION

Enzyme preparations - sources of β-galactosidase, are widely used in the dairy industry. However, there is no standardized methods to determine their activity. Existing methods differ by the type of substrate used, conditions of analysis and how to define the products of the hydrolysis. The aim of this work is to study the process of conversion biocatalytic milk sugar and experimental substantiation of methods of analysis of hydrolysis products, providing reliable results determine activity of β-galactosidase. Found that of chemical methods glucose oxidase showed a greater sensitivity towards low concentrations of the product of hydrolysis is glucose formed in the conversion process lactose. The results of testing showed that the mass concentration of the glucose formed in hydrolysis of lactose, HPLC tested, consistent with data obtained using the glucose oxidase method. Set optimum range, which was observed directly proportional to the level of glucose concentration rise in reaction mixture from the lengthy process and the concentration of enzyme: hydrolysis time - from 10 to 20 min, produced glucose - from 20 µcg to 40 mcg, while the mass of enzyme - 25.0·10-6, and from 40 mcg to 80 mcg, when mass -5,0·10-5 g. When the values of absorbance of the reaction mixture ranged 0,045-0,300 to ensure reliable performance. Studies on determination of the activity of the enzyme preparations confirmed received experimental data and allowed to choose optimum parameters and conditions analysis for introducing them to the method for determining the activity β-galactosidase using glucose oxidase reagent: time of inactivation enzyme (control) -10 min. in a boiling water bath, substrate - 2% solution of lactose, pH of the substrate-6,0, hydrolysis time 15 min. at 30° c.