New alkalophilic β-galactosidase with high activity in alkaline pH region from Teratosphaeria acidotherma AIU BGA-1

Yamada, Miwa; Chiba, Serina; Endo, Yuta; Isobe, Kimiyasu

doi:10.1016/j.jbiosc.2016.07.003

Cited by 10 publications

(5 citation statements)

References 20 publications

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“…Surprisingly, the inhibition of enzymatic activity by EDTA was clearly recovered in the presence of 1.0 mM Mn 2+ (data not shown). These results clearly indicate that the enzyme having a metallic nature which Mn 2+ play an important role for activating and protecting the active site of the enzyme against inhibitors [3]. Similarly, the recombinant β-galactosidase gene (PbBGal2A) from Paenibacillus barengoltzii CAU904 was reported as a metal-dependent enzyme [35], which was strongly inactivated by EDTA and highly stimulated by the presence of each of K + , Na + ,Mn 2+ , and Mg 2+ .…”

Section: Effect Of Ph Temperature and Salinity On β-Galactosidasementioning

confidence: 81%

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Activation of LacZ gene in Escherichia coli DH5α via α-complementation mechanism for β-galactosidase production and its biochemical characterizations

Hamed

Khedr

Abdelraof

2020

Journal of Genetic Engineering and Biotechnology

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Background Plasmid propagation in recombination strains such as Escherichia coli DH5α is regarded as a beneficial instrument for stable amplification of the DNA materials. Here, we show trans-conjugation of pGEM-T cloning vector (modified Promega PCR product cloning vector with tra genes, transposable element (Tn5)) and M13 sequence via α-complementation mechanism in order to activate β-d-galactosidase gene in DH5α strain (non-lactose-fermenting host). Results Trans-conjugation with pGEM-T allows correction of LacZ gene deletion through Tn5, and successful trans-conjugants in DH5α host cells can be able to produce active enzyme, thus described as lactose fermenting strain. The intracellular β-galactosidase was subjected to precipitation by ammonium sulfate and subsequently gel filtration, and the purified enzyme showed a molecular weight of approximately 72-kDa sodium dodecyl sulfate-polyacrylamid gel electrophoresis. The purified enzyme activity showed an optimal pH and temperature of 7.5 and 40 °C, respectively; it had a high stability within pH 6–8.5 and moderate thermal stability up to 50 °C. Conclusion Trans-conjugant of E. coli DH5α- lacZ∆M15 was successfully implemented. UV mutagenesis of the potent trans-conjugant isolate provides an improvement of the enzyme productivity. The enzymatic competitive inhibition by d-galactose and hydrolysis of lactose at ambient temperatures could make this enzyme a promising candidate for use in the dairy industry.

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Section: Effect Of Ph Temperature and Salinity On β-Galactosidasementioning

confidence: 81%

“…β-Galactosidase or lactase (EC 3.2.1.23) is the enzyme that catalyzes the conversion of lactose to monosaccharide sugars. People with lactose intolerance are unable to make enough lactase enzymes which in turn causes the inability to consume dairy products [1][2][3][4].…”

Section: Introductionmentioning

confidence: 99%

Activation of LacZ gene in Escherichia coli DH5α via α-complementation mechanism for β-galactosidase production and its biochemical characterizations

Hamed

Khedr

Abdelraof

2020

Journal of Genetic Engineering and Biotechnology

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“…and consequently, the production of β‐galactosidase, which can be explained through the presence of galactans (Husain 2010) and magnesium, an indispensable mineral for the enzyme activity (Yamada et al . 2017).…”

Section: Discussionmentioning

confidence: 99%

“…1996; Yamada et al . 2017). The use of tap water as moistening is interesting because of its availability and low cost.…”

Section: Discussionmentioning

confidence: 99%

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Production of β‐galactosidase by Trichoderma sp. through solid‐state fermentation targeting the recovery of galactooligosaccharides from whey cheese

Jesus

Guimarães

2020

J. Appl. Microbiol.

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Aims Optimization of β‐galactosidase production by Trichoderma sp. under solid‐state fermentation using wheat bran as solid substrate through an experimental design and its application targeting the recovery of galactooligosaccharides (GOS) from whey cheese. Methods and Results The β‐galactosidase production by Trichoderma sp. increased 2·3‐fold (2·67 U g−1 of substrate) culturing the fungus at 30°C for 187 h, at an inoculum of 105 spores per ml, and a 1 : 1·65 (w/v) ratio of wheat bran to tap water. The best enzyme activity was obtained at 55°C and pH 4·5. The catalytic activity was maintained for up to 180 min incubating at 35–45°C, and above 50% at acidic or alkaline pH for up to 24 h. It also presented resistance to chemical compounds. β‐galactosidase catalysed the hydrolysis of the lactose and the transgalactosylation reaction leading to the production of GOS. Conclusion Trichoderma sp. produced β‐galactosidase with transgalactosylation activity that may be used to recover GOS, products with high added value, from whey cheese. Significance and Impact of the Study β‐galactosidases are used in different industrial sectors. Therefore, the Trichoderma β‐galactosidase is a promising alternative for the production of GOS as prebiotic from the dairy effluents, contributing to the reduction in the environmental impact.

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β-Galactosidases from an Acidophilic Fungus, Teratosphaeria acidotherma AIU BGA-1

Isobe

Yamada

2019

Fungi in Extreme Environments: Ecological Role and Biotechnological Significance

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New alkalophilic β-galactosidase with high activity in alkaline pH region from Teratosphaeria acidotherma AIU BGA-1

Cited by 10 publications

References 20 publications

Activation of LacZ gene in Escherichia coli DH5α via α-complementation mechanism for β-galactosidase production and its biochemical characterizations

Activation of LacZ gene in Escherichia coli DH5α via α-complementation mechanism for β-galactosidase production and its biochemical characterizations

Production of β‐galactosidase by Trichoderma sp. through solid‐state fermentation targeting the recovery of galactooligosaccharides from whey cheese

β-Galactosidases from an Acidophilic Fungus, Teratosphaeria acidotherma AIU BGA-1

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