Functional Characterization of Recombinant Endo-Levanase (LevBk) from Bacillus koreensis HL12 on Short-Chain Levan-Type Fructooligosaccharides Production

IF 1.9 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

The Protein Journal Pub Date : 2022-08-06 DOI:10.1007/s10930-022-10069-2

Hataikarn Lekakarn, Benjarat Bunterngsook, Phuphiphat Jaikaew, Thanyanun Kuantum, Rungtiva Wansuksri, Verawat Champreda

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引用次数: 1

Abstract

Levan-type fructooligosaccharides (L-FOSs) are a prominent class of non-digestible oligosaccharides with potential as nutritional prebiotics. Endo-levanase, which randomly hydrolyzes β-(2,6)-linkages in fructans, is a promising enzyme for short-chain FOS production. In this work, a recombinant levanase (LevBk) from Bacillus koreensis strain HL12 was characterized. Soluble LevBk protein was produced in Escherichia coli BL21(DE3) system at 40 mg/L of culture medium. Based on sequence and structural analysis, LevBk was classified as a member of endo-levanase in GH32 family containing N-terminal substrate binding pocket and C-terminal β-sandwich domains. LevBk optimally worked at 45 °C, pH 6.0 with the specific activity of 2.43 U/mg. Based on enzymatic hydrolysis, short-chain L-FOSs with degree of polymerization (DP) of 2–4 were produced from hydrolysis of timothy grass levan under optimal conditions for 9–24 h. With its ability to produce L-FOSs with specific chain lengths, LevBk could be attractively applied for converting of levan containing material to high value-added sweetener in the biorefinery industry.

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韩国芽孢杆菌HL12重组内切利末酸酶(levane - enase, LevBk)在短链利末型低聚果糖生产中的功能研究

利凡型低聚果糖(L-FOSs)是一类重要的不可消化低聚糖，具有作为营养益生元的潜力。内旋左旋酶可以随机水解果聚糖中的β-(2,6)键，是一种很有前途的短链果寡糖酶。本研究对韩国芽孢杆菌HL12菌株的重组乙酰化酶(LevBk)进行了鉴定。在大肠杆菌BL21(DE3)体系中，在40 mg/L培养基中产生可溶性的LevBk蛋白。根据序列和结构分析，LevBk属于GH32家族内切乙酰化酶成员，含有n端底物结合口袋和c端β-三明治结构域。LevBk在45°C、pH 6.0条件下的最佳活性为2.43 U/mg。通过酶解，在最佳条件下水解9 ~ 24 h，得到了聚合度(DP)为2 ~ 4的短链L-FOSs。LevBk具有生产特定长度链的L-FOSs的能力，可以在生物炼制工业中将含levan的物质转化为高附加值的甜味剂。

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来源期刊

The Protein Journal 生物-生化与分子生物学

CiteScore

5.20

自引率

0.00%

发文量

审稿时长

12 months

期刊介绍： The Protein Journal (formerly the Journal of Protein Chemistry) publishes original research work on all aspects of proteins and peptides. These include studies concerned with covalent or three-dimensional structure determination (X-ray, NMR, cryoEM, EPR/ESR, optical methods, etc.), computational aspects of protein structure and function, protein folding and misfolding, assembly, genetics, evolution, proteomics, molecular biology, protein engineering, protein nanotechnology, protein purification and analysis and peptide synthesis, as well as the elucidation and interpretation of the molecular bases of biological activities of proteins and peptides. We accept original research papers, reviews, mini-reviews, hypotheses, opinion papers, and letters to the editor.