来源于生孢噬纤维菌的新型β-葡萄糖苷酶SmBgl3A的性质研究

doi:10.3969/j.issn.1673-1689.2023.01.007

首页 > 过刊浏览>2023年第42卷第1期 >83-92. DOI:10.3969/j.issn.1673-1689.2023.01.007

来源于生孢噬纤维菌的新型β-葡萄糖苷酶SmBgl3A的性质研究
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                        10.3969/j.issn.1673-1689.2023.01.007
                    
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Characterization of A Novel β-glucosidase SmBgl3A from Sporocytophaga sp. CX11

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通过表达水平分析比较，从生孢噬纤维菌Sporocytophaga sp. CX11基因组中筛选出在菌株降解纤维二糖过程中起关键作用的β-葡萄糖苷酶基因SmBgl3A。该基因含有2 283 bp碱基对，编码760个氨基酸，其编码蛋白质与来源于Bacteroides ovatus的β-葡萄糖苷酶BoGH3B同源性为46.76 %。将该基因在大肠杆菌中进行异源表达，粗酶液经Ni-NTA亲和层析一步纯化得到电泳级纯度的酶蛋白SmBgl3A，重组蛋白质相对分子质量与理论值（81.2×10⁴）一致。酶学性质表征结果表明，SmBgl3A最适温度和pH分别为35 ℃和6.5；pH在5.0~7.0、温度在40 ℃以下时，稳定性较好，同时有着较好的NaCl耐受性。SmBgl3A的最适底物为pNPG，以pNPG为底物时的比活力为14.74 U/mg；K_m为10.88 mmol/L，k_cat为46.72 s< sup > -1。SmBgl3A作为一种新型的、酶学性质优良的β-葡萄糖苷酶，进一步扩展了β-葡萄糖苷酶的种类，为该酶在纤维素降解、食品工业等领域的应用奠定基础。

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A β-glucosidase gene SmBgl3A, which might play a key role in cellobiose degradation and utilization, was screened from Sporocytophaga sp. CX11 through the analysis and comparison of expression level. The gene contains 2 283 bp and encodes 760 amino acids. The encoded protein of SmBgl3A has 46.76% homology with the β-glucosidase BoGH3B from Bacteroides ovatus. The gene SmBgl3A was heterologously expressed in Escherichia coli. The electrophoretically pure recombinant protein SmBgl3A was achieved by the purification of crude enzymes using Ni-NTA affinity chromatography, and the relative molecular weight of recombinant protein was consistent with the theoretical value (81.2×10⁴). Characterization of enzymatic properties showed that the optimum temperature and pH of SmBgl3A were 35 ℃ and 6.5, respectively. When the pH value was 5.0~7.0 and the temperature was below 40 ℃, SmBgl3A was stable. In addition, SmBgl3A showed good NaCl tolerance. The optimal substrate for SmBgl3A was pNPG, and the specific enzyme activity with pNPG as substrate was 14.74 U/mg. K_m and k_cat of SmBgl3A towards pNPG were 10.88 mmol/L and 46.72 s^-1, respectively. SmBgl3A is a novel enzyme with excellent enzymatic properties, further expanding the kinds of β-glucosidase and laying a foundation for the enzyme application in cellulose degradation, food industry and other fields.

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费嵩禹,王梓懿,荆晓凤,杨帆,陈晓艺,李宪臻.来源于生孢噬纤维菌的新型β-葡萄糖苷酶SmBgl3A的性质研究[J].食品与生物技术学报,2023,42(1):83-92.

FEI Songyu, WANG Ziyi, JING Xiaofeng, YANG Fan, CHEN Xiaoyi, LI Xianzhen. Characterization of A Novel β-glucosidase SmBgl3A from Sporocytophaga sp. CX11[J]. Journal of Food Science and Biotechnology,2023,42(1):83-92.

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在线发布日期: 2023-02-06
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