Expression and characterization of β-galactosidase from fecal microbes of Rhinopithecus bieti
Author:
  • Article
  • | |
  • Metrics
  • |
  • Reference [33]
  • |
  • Related
  • | | |
  • Comments
    Abstract:

    [Objective] The study was to express and characterize β-galactosidase from the fecal microbes of Rhinopithecus bieti. [Methods] A β-galactosidase gene galRBM20_1 was cloned from the fecal microbial metagenome of Rhinopithecus bieti. The pEASY-E2-galRBM20_1 plasmid was constructed and transformed into Escherichia coli BL21 (DE3). The expression was induced by Isopropyl β-D-1-thiogalactopyranoside and the enzymatic properties were studied.[Results] The optimum pH of β-galactosidase galRBM20_1 was 5.0, and the enzyme retained more than 70% activity between pH 4 and 7. The optimum temperature is 45℃ and the enzyme was stable at 37℃ and 45℃, retaining nearly 100% activity after incubation for 1 h. In addition, the enzyme had strong NaCl stability. After 1 h of 1 to 5 mol/L NaCl treatment, the relative enzyme activity maintained 100%; when the concentration of NaCl was 4 mol/L, the activity of β-galactosidase galRBM20_1 was the highest (146%); it still had activity after treatment with 2.5 mol/L NaCl at 45℃ for 24 h. [Conclusion] β-galactosidase galRBM20_1 has good salt tolerance and wide pH range.

    Reference
    [1] Vasileva N, Ivanov Y, Damyanova S, Kostova I, Godjevargova T. Hydrolysis of whey lactose by immobilized β-galactosidase in a bioreactor with a spirally wound membrane. International Journal of Biological Macromolecules, 2016, 82:339-346.
    [2] Pereira-Rodriguez Á, Fernández-Leiro R, González-Siso MI, Cerdán ME, Becerra M, Sanz-Aparicio J. Structural basis of specificity in tetrameric Kluyveromyces lactis β-galactosidase. Journal of Structural Biology, 2012, 177(2):392-401.
    [3] Grosová Z, Rosenberg M, Rebroš M. Perspectives and applications of immobilised β-galactosidase in food industry-a review. Czech Journal of Food Sciences, 2008, 26(1):1-14.
    [4] Sharma SK, Li SH, Micic M, Orbulescu J, Weissbart D, Nakahara H, Shibata O, Leblanc RM. β-galactosidase Langmuir monolayer at air/X-gal subphase interface. The Journal of Physical Chemistry B, 2016, 120(48):12279-12286.
    [5] Chanalia P, Gandhi D, Attri P, Dhanda S. Purification and characterization of β-galactosidase from probiotic Pediococcus acidilactici and its use in milk lactose hydrolysis and galactooligosaccharide synthesis. Bioorganic Chemistry, 2018, 77:176-189.
    [6] Felicilda-Reynaldo RFD, Kenneally M. Digestive enzyme replacement therapy:pancreatic enzymes and lactase. Medsurg Nursing, 2016, 25(3):182-185.
    [7] Harju M, Kallioinen H, Tossavainen O. Lactose hydrolysis and other conversions in dairy products:technological aspects. International Dairy Journal, 2012, 22(2):104-109.
    [8] Saqib S, Akram A, Halim SA, Tassaduq R. Sources of β-galactosidase and its applications in food industry. 3 Biotech, 2017, 7(1):79.
    [9] Juajun O, Nguyen TH, Maischberger T, Iqbal S, Haltrich D, Yamabhai M. Cloning, purification, and characterization of β-galactosidase from Bacillus licheniformis DSM 13. Applied Microbiology and Biotechnology, 2011, 89:645-654.
    [10] Cheng JJ, Romantsov T, Engel K, Doxey AC, Rose DR, Neufeld JD, Charles TC. Functional metagenomics reveals novel β-galactosidases not predictable from gene sequences. PLoS One, 2017, 12(3):e0172545.
    [11] Gans J, Wolinsky M, Dunbar J. Computational improvements reveal great bacterial diversity and high metal toxicity in soil. Science, 2005, 309(5739):1387-1390.
    [12] Wei DZ, Chen SX, Wang XL, Yuan QS, Yu JT. Properties of β-galactosidase from Bacillus Stearothermophilus. Microbiology China, 2001, 28(1):18-22. (in Chinese)魏东芝, 陈少欣, 王筱兰, 袁勤生, 俞俊棠. 嗜热脂肪芽孢杆菌β-半乳糖苷酶的性质. 微生物学通报, 2001, 28(1):18-22.
    [13] Lorenz P, Eck J. Screening for novel industrial biocatalysts. Engineering in Life Sciences, 2004, 4(6):501-504.
    [14] Matsuzawa T, Yaoi K. Screening, identification, and characterization of a novel saccharide-stimulated β-glycosidase from a soil metagenomic library. Applied Microbiology and Biotechnology, 2017, 101(2):633-646.
    [15] Wierzbicka-Woś A, Bartasun P, Cieśliński H, Kur J. Cloning and characterization of a novel cold-active glycoside hydrolase family 1 enzyme with β-glucosidase, β-fucosidase and β-galactosidase activities. BMC Biotechnology, 2013, 13:22.
    [16] Deng J, Zhang Q, Wei YL, Lin LB. A β-galactosidase derived from hot springs metagenome and its characteristics. Food and Fermentation Industries, 2014, 40(5):60-64. (in Chinese)邓建, 张琦, 魏云林, 林连兵. 来源于热泉宏基因组的β-半乳糖苷酶及其特征. 食品与发酵工业, 2014, 40(5):60-64.
    [17] Maruthamuthu M, Van Elsas JD. Molecular cloning, expression, and characterization of four novel thermo-alkaliphilic enzymes retrieved from a metagenomic library. Biotechnology for Biofuels, 2017, 10:142.
    [18] Xu B, Xu WJ, Li JJ, Dai LM, Xiong CY, Tang XH, Yang YJ, Mu YL, Zhou JP, Ding JM, Wu Q, Huang ZX. Metagenomic analysis of the Rhinopithecus bieti fecal microbiome reveals a broad diversity of bacterial and glycoside hydrolase profiles related to lignocellulose degradation. BMC Genomics, 2015, 16:174.
    [19] Dai LM, Deng M, Huang ZX, Wang YJ, Li JJ, Zhou JP, Mu YL, Xu B. Gene diversity of the glycosyl hydrolase family 10 xylanase in the fecal microorganism of Rhinopithecus bieti. Microbiology China, 2016, 43(1):44-50. (in Chinese)戴利铭, 邓梦, 黄遵锡, 王余娇, 李俊俊, 周峻沛, 穆跃林, 许波. 滇金丝猴粪便微生物GH10家族木聚糖酶基因多样性. 微生物学通报, 2016, 43(1):44-50.
    [20] Xu B, Dai LM, Li JJ, Deng M, Miao HB, Zhou JP, Mu YL, Wu Q, Tang XH, Yang YJ, Ding JM, Han NY, Huang ZX. Molecular and biochemical characterization of a novel xylanase from Massilia sp. RBM26 isolated from the feces of Rhinopithecus bieti. Journal of Microbiology and Biotechnology, 2016, 26(1):9-19.
    [21] 戴利铭. 滇金丝猴粪便微生物木聚糖降解酶研究. 云南师范大学生命科学学院硕士学位论文, 2016.
    [22] Sutendra G, Wong S, Fraser ME, Huber RE. β-galactosidase (Escherichia coli) has a second catalytically important Mg2+ site. Biochemical and Biophysical Research Communications, 2007, 352(2):566-570.
    [23] Liu Y, Chen Z, Jiang ZQ, Yan QJ, Yang SQ. Biochemical characterization of a novel β-galactosidase from Paenibacillus barengoltzii suitable for lactose hydrolysis and galactooligosaccharides synthesis. International Journal of Biological Macromolecules, 2017, 104:1055-1063.
    [24] Wang GX, Gao Y, Hu B, Lu XL, Liu XY, Jiao BH. A novel cold-adapted β-galactosidase isolated from Halomonas sp. S62:gene cloning, purification and enzymatic characterization. World Journal of Microbiology and Biotechnology, 2013, 29(8):1473-1480.
    [25] Cardoso BB, Silvério SC, Abrunhosa L, Teixeira JA, Rodrigues LR. β-galactosidase from Aspergillus lacticoffeatus:a promising biocatalyst for the synthesis of novel prebiotics. International Journal of Food Microbiology, 2017, 257:67-74.
    [26] Wang SD, Guo GS, Li L, Cao LC, Tong L, Ren GH, Liu YH. Identification and characterization of an unusual glycosyltransferase-like enzyme with β-galactosidase activity from a soil metagenomic library. Enzyme and Microbial Technology, 2014, 57:26-35.
    [27] Guo BS, Zheng F, Crouch L, Cai ZP, Wang M, Bolam DN, Liu L, Voglmeir J. Cloning, purification and biochemical characterisation of a GH35 beta-1,3/beta-1,6-galactosidase from the mucin-degrading gut bacterium Akkermansia muciniphila. Glycoconjugate Journal, 2018, 35(3):255-263.
    [28] Wang K, Li G, Yu SQ, Zhang CT, Liu YH. A novel metagenome-derived β-galactosidase:gene cloning, overexpression, purification and characterization. Applied Microbiology and Biotechnology, 2010, 88(1):155-165.
    [29] Sheridan PP, Brenchley JE. Characterization of a salt-tolerant family 42β-galactosidase from a psychrophilic antarctic Planococcus isolate. Applied and Environmental Microbiology, 2000, 66(6):2438-2444.
    [30] Rabausch U, Juergensen J, Ilmberger N, Böhnke S, Fischer S, Schubach B, Schulte M, Streit WR. Functional screening of metagenome and genome libraries for detection of novel flavonoid-modifying enzymes. Applied and Environmental Microbiology, 2013, 79(15):4551-4563.
    [31] Simon C, Daniel R. Achievements and new knowledge unraveled by metagenomic approaches. Applied Microbiology and Biotechnology, 2009, 85(2):265-276.
    [32] Iqbal HA, Feng ZY, Brady SF. Biocatalysts and small molecule products from metagenomic studies. Current Opinion in Chemical Biology, 2012, 16(1/2):109-116.
    [33] Schmeisser C, Steele H, Streit WR. Metagenomics, biotechnology with non-culturable microbes. Applied Microbiology and Biotechnology, 2007, 75(5):955-962.
    Related
    Cited by
Get Citation

Wenhong Zhang, Yanxia Yang, Zhengfeng Yang, Zunxi Huang, Junjun Li, Xianghua Tang, Yunjuan Yang, Qian Wu, Yuelin Mu, Nanyu Han, Bo Xu. Expression and characterization of β-galactosidase from fecal microbes of Rhinopithecus bieti. [J]. Acta Microbiologica Sinica, 2019, 59(8): 1561-1575

Copy
Share
Article Metrics
  • Abstract:936
  • PDF: 1230
  • HTML: 1320
  • Cited by: 0
History
  • Received:October 19,2018
  • Revised:January 24,2019
  • Online: July 31,2019
Article QR Code