Overproduction of thermostable β-galactosidase in Escherichia coli, its purification and molecular structure

Saito, Takao; Kato, Katsuya; Maeda, Sumio; Suzuki, Tohru; Shiba, Sumihisa; Iijima, Shinji; Kobayashi, Takeshi

doi:10.1016/0922-338x(92)90260-2

Cited by 9 publications

(4 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The relative initial rate of hydrolysis of PNP glycoside is expressed as a percentage of that obtained with ONPG. 12,14,18,22,24,25,27,28,31). The thermostability of ␤-galactosidase from S. elviae was slightly lower at 85ЊC than those from Sulfolobus solfataricus (22) and Thermotoga maritima (4).…”

Section: Discussionmentioning

confidence: 95%

“…As the S. elviae ␤-galactosidase retained 90% of its activity after preincubation at 80ЊC, it is comparable with ␤-galactosidases from Thermus aquaticus (31), Bacillus stearothermophilus (7), Bacillus acidocaldarius (12), Thermoaerobacter sp. (14), Saccharopolyspora rectivirgula (18), and the thermophilic anaerobe strain NA10 (24). Almost all the thermostable ␤-galactosidases have so far been found in thermophilic and thermotolerant microorganisms.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Purification and properties of a novel thermostable galacto-oligosaccharide-producing beta-galactosidase from Sterigmatomyces elviae CBS8119

Onishi

Tanaka

1995

Appl Environ Microbiol

View full text Add to dashboard Cite

A thermostable ␤-galactosidase which catalyzed the production of galacto-oligosaccharide from lactose was solubilized from a cell wall preparation of Sterigmatomyces elviae CBS8119. The enzyme was purified to homogeneity by means of chromatography on DEAE-Toyopearl, Butyl-Toyopearl, Chromatofocusing, and p-aminobenzyl 1-thio-␤-D-galactopyranoside agarose columns. The molecular weight of the purified enzyme was estimated to be about 170,000 by gel filtration with a Highload-Superdex 200pg column and 86,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its isoelectric point, determined by polyacrylamide gel electrofocusing, was 4.1. The optimal temperature for enzyme activity was 85؇C. It was stable at temperatures up to 80؇C for 1 h. The optimal pH range for the enzyme was 4.5 to 5.0, it was stable at pH 2.5 to 7.0, and its activity was inhibited by Hg 2؉. The K m values for o-nitrophenyl-␤-D-galactopyranoside and lactose were 9.5 and 2.4 mM, respectively, and the maximum velocities for these substrates were 96 and 240 mol/min per mg of protein, respectively. In addition, this enzyme possessed a high level of transgalactosylation activity. Galacto-oligosaccharides, including tri-and tetrasaccharides, were produced with a yield, by weight, of 39% from 200-mg/ml lactose.

show abstract

Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Purification and properties of a novel thermostable galacto-oligosaccharide-producing beta-galactosidase from Sterigmatomyces elviae CBS8119

Onishi

Tanaka

1995

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…Nonetheless, the protein production could even be higher by using, for example, a fed-bacth culture (Saito et al, 1992). It has to be considered that the time required to grow the E. coli transformant (16 h at 37°C) is much shorter than that of the wild type B. subtilis (7 days at 10°C).…”

Section: Discussionmentioning

confidence: 99%

Cloning and expression of ?-galactosidase from psychotrophic Bacillus subtilis KL88 into Escherichia coli

Torres

Lee

1995

Biotechnol Lett

View full text Add to dashboard Cite

The B-galactosidase gene from Bacillus subtilis KL88 was cloned into Escherichia coli and the gene product characterized for its potential use in the dairy industry. The two recombinant plasmids that we obtanied encoded a B-galactosidase with the same catalytic and thermal characteristics as the native B-galactosidase from B. subtilis. The recombinant B-galactosidases exhibited high activity at low temperature (10°C), with maximum activity at 50°C and an optimum pH of 6.0. Its molecular weight was estimated to be 90 Kd. The restriction maps of the recombinant plasmids were constructed. The 13-galactosidase gene was located in a 2.3 Kb fragment.

show abstract

“…Nakayama et al, , 1993Nakao et al, 1994;20. Sienkiewicz and Riedel, 1990, p256;Saito et al, 1992. complex and responds to ionic concentrations of a range of elements, namely, K, Mn and Mg. The enzymes are inhibited by Na + and Ca 2+ .…”

Section: Investigations Of the Major Transgalactosylation Enzyme Sourcesmentioning

confidence: 99%

Galacto-oligosaccharides and Other Products Derived from Lactose

Playne¹,

Crittenden

2009

Advanced Dairy Chemistry

View full text Add to dashboard Cite

Lactose is the precursor for a number of compounds derived by chemical, physical or enzymatic conversion that have an established and expanding place in the health and food industries. These include lactulose and galactooligosaccharides both of which are manufactured in large tonnages worldwide. Lactitol and lactosucrose are produced commercially, but in much smaller amounts, while lactobionic acid is produced for limited industrial and medical applications and for research use. The only other lactose derivative of commercial interest is an isomer of galactose called tagatose in which there is an emerging interest as a sweetener.None of these compounds occur naturally in cow's milk except in trace amounts. Yet, lactose-derived oligosaccharides occur naturally in human milk at relatively high concentrations (5-12 g/L) and play a major role in directing the development of the intestinal microbiota of infants. One of the major and emerging uses of lactose derivatives has been to emulate the physiological effects of these natural oligosaccharides, and -galacto-oligosaccharides (GOSs) and lactulose in particular have found applications as prebiotics in functional foods and beverages. GOSs are also classified nowadays as dietary fibre. Prebiotic -galacto-oligosaccharides are also available commercially, but since they are produced typically from soybean, they are not discussed here. Lactulose is an important pharmaceutical used in the treatment of constipation ,Springer ScienceþBusiness Media, LLC 2009 121 BIFIDUS GROWTH FACTOR 2 Casein peptides (non-glycosylated) (after hydrolysis using a proteinase) BIFIDUS GROWTH FACTOR 3 Pantethine OLIGOSACCHARIDES Stachyose Fructo-oligosaccharides Galacto-oligosaccharides Iso-malto oligosaccharides Xylo oligosaccharides DI-AND TRI-SACCHARIDES Raffinose Lactulose Lactosucrose CELL-FREE EXTRACTS Propionibacterium freudenreichii LACTOFERRIN Fe, Cu and Zn complexes Galacto-oligosaccharides and Other Products Derived from Lactose 125

show abstract

Overproduction of thermostable β-galactosidase in Escherichia coli, its purification and molecular structure

Cited by 9 publications

References 17 publications

Purification and properties of a novel thermostable galacto-oligosaccharide-producing beta-galactosidase from Sterigmatomyces elviae CBS8119

Purification and properties of a novel thermostable galacto-oligosaccharide-producing beta-galactosidase from Sterigmatomyces elviae CBS8119

Cloning and expression of ?-galactosidase from psychotrophic Bacillus subtilis KL88 into Escherichia coli

Galacto-oligosaccharides and Other Products Derived from Lactose

Contact Info

Product

Resources

About