Purification and Characterization of a Novel β-Galactosidase From the Thermoacidophile Alicyclobacillus vulcanalis

Murphy, J. Brad; Ryan, Michael P.; Walsh, Gary

doi:10.1007/s12010-020-03233-w

Cited by 17 publications

(8 citation statements)

References 50 publications

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“…barengoltzii (pH 7.5) but higher than that of the β-galactosidase from Bacillus aryabhattai GEL-09 (pH 6.0) . TsGal48 showed good pH stability in the range of pH 5–8 (Figure b), which is wider than that of some bacterial β-galactosidases from thermoacidophile Alicyclobacillus vulcanalis (pH 5.5–6.5), P. barengoltzii (pH 6.0–8.0), and B.…”

Section: Results and Discussionmentioning

confidence: 94%

“…The technology was more efficient than active charcoal column separation, which is regarded as green and sustainable due to its low cost, easy operation, and low energy consumption. , Moreover, the technology has first been applied to separate LNnT to date, and it is suitable for a large-scale production . In addition, high reaction temperature (70 °C) and short reaction time (1 h) for LNnT synthesis can reduce the risk of microbial contamination in industrial production . Moreover, the yield of LNnT is 2 folds higher than that of the previous system …”

Section: Results and Discussionmentioning

confidence: 99%

“…29 Moreover, TsGal48 was stable up to 75 °C (Figure 2d), which is higher than those of many β-galactosidases from Microvirga sp. strain MC18, 26 B. aryabhattai GEL-09, 27 and A. vulcanalis 28 but lower than that of Tnap1577 from T. naphthophila RUK-10. 29 Most of the β-galactosidases exhibited strict β-galactosidase activity, such as β-galactosidases from P. barengoltzii and deep-sea bacterium Alteromonas sp.…”

Section: Sequence Analysis Of Tsgal48mentioning

confidence: 86%

“…12 The optimal pH of TsGal48 was pH 6.5 (Figure 2a), which is lower than that of the β-galactosidase from P. barengoltzii (pH 7.5) 12 but higher than that of the β-galactosidase from Bacillus aryabhattai GEL-09 (pH 6.0). 27 TsGal48 showed good pH stability in the range of pH 5−8 (Figure 2b), which is wider than that of some bacterial β-galactosidases from thermoacidophile Alicyclobacillus vulcanalis (pH 5.5−6.5), 28 P. barengoltzii (pH 6.0−8.0), 12 and B. aryabhattai GEL-09 (pH 5.0− 7.0). 27 The optimal temperature of TsGal48 was up to 70 °C (Figure 2c), which is higher than that of some β-galactosidases from Microvirga sp.…”

Section: Sequence Analysis Of Tsgal48mentioning

confidence: 96%

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Protein Engineering of a Novel β-Galactosidase from Thermus scotoductus for Efficient Synthesis of Lacto-N-Neotetraose from Chitin Powder

Wang,

Xiang,

Liu

et al. 2024

J. Agric. Food Chem.

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A novel β-galactosidase (TsGal48) from Thermus scotoductus was cloned, and the enzyme was biochemically characterized. TsGal48 catalyzed the synthesis of lacto-N-neotetraose (LNnT) from lactose via the transglycosylation reaction with a maximal yield of 20%, which is the highest yield for the synthesis of LNnT so far. To further improve the yield of LNnT, TsGal48 was successfully engineered by directed evolution and site-saturation mutagenesis. A mutated β-galactosidase (mTsGal48) was selected and characterized. mTsGal48 produced LNnT with a yield of 27.7 g/L, which is 1.4-fold higher than that of TsGal48 (19.7 g/L). Then, a developed strategy for LNnT synthesis from chitin powder was provided in a 30 L bioreactor. The reaction process included chitin powder hydrolysis, lacto-N-triose II (LNT2) synthesis, and LNnT synthesis. The reaction time was reduced from 44 to 17 h in chitin powder hydrolysis and LNT2 synthesis. The content of LNnT was up to 25 g/L in the multienzyme system. The green and efficient route may be suitable for large-scale production of LNnT from chitin powder.

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Section: Results and Discussionmentioning

confidence: 94%

Section: Results and Discussionmentioning

confidence: 99%

Section: Sequence Analysis Of Tsgal48mentioning

confidence: 86%

Section: Sequence Analysis Of Tsgal48mentioning

confidence: 96%

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Protein Engineering of a Novel β-Galactosidase from Thermus scotoductus for Efficient Synthesis of Lacto-N-Neotetraose from Chitin Powder

Wang,

Xiang,

Liu

et al. 2024

J. Agric. Food Chem.

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“…However, the recombinant expression of β-galactosidases in E. coli, Lactobacillus, and Lactococcus are usually cytoplasmic, making the purification tedious and costly [ 12 ]. Besides heterogeneous expression, many of β-galactosidases were produced by native fermentation such as Alicyclobacillus vulcanalis [ 30 ], Kluyveromyces lactis De [ 31 ] and Aspergillus terreus [ 2 ]. However, some typical problems exist during their production, e.g., the fermentation may contain other endogenous proteins leading to complex downstream processing and high economic costs.…”

Section: Resultsmentioning

confidence: 99%

β-galactosidase GALA fromBacillus circulanswith high transgalactosylation activity

Yan

Guan

et al. 2021

Bioengineered

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β-galactosidase catalyzes lactose hydrolysis and transfers reactions to produce prebiotics such as galacto-oligosaccharides (GOS) with potential application in the food industry and pharmaceuticals. However, there is still a need for improved transgalactosylation activity of β-galactosidases and reaction conditions of GOS production in order to maximize GOS output and reduce production costs. In this study, a β-galactosidase gene, galA, from Bacillus circulans was expressed in Pichia pastoris, which not only hydrolyzed lactose but also had strong transgalactosylation activity to produce GOS. Response surface methodology was adopted to investigate the effects of temperature, enzyme concentration, pH, initial lactose concentration and reaction time on production of GOS and optimize the reaction conditions for GOS.The optimal pH for the enzyme was 6.0 and remained stable in neutral and basic conditions. Meanwhile, GALA showed most activity at 50℃ and retained considerable activity at lower temperature 30-40°C, indicating this enzyme could work under mild conditions. The enzyme concentration and temperature were found to be the critical parameters affecting the transgalactosylation activity. Response surface methodology showed that the optimal enzyme concentration, initial lactose concentration, temperature, pH, and reaction time were 3.03 U/mL, 500 g/L, 30℃, 5.08, and 4 h, respectively. Under such conditions, the maximum yield of GOS was 252.8 g/L, accounting for approximately 50.56% of total sugar. This yield can be considered relatively high compared to those obtained from other sources of β-galactosidases, implying a great potential for GALA in the industrial production and application of GOS.

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Immobilization of β-Galactosidase in Calcium Alginate Beads

Martins

Figueira

Castilho

2021

Basic Protocols in Encapsulation of Food Ingredients

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Purification and Characterization of a Novel β-Galactosidase From the Thermoacidophile Alicyclobacillus vulcanalis

Cited by 17 publications

References 50 publications

Protein Engineering of a Novel β-Galactosidase from Thermus scotoductus for Efficient Synthesis of Lacto-N-Neotetraose from Chitin Powder

Protein Engineering of a Novel β-Galactosidase from Thermus scotoductus for Efficient Synthesis of Lacto-N-Neotetraose from Chitin Powder

β-galactosidase GALA fromBacillus circulanswith high transgalactosylation activity

Immobilization of β-Galactosidase in Calcium Alginate Beads

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