Efficient production of a highly active lysozyme from European flat oyster Ostrea edulis

IF 4.1 2区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Bo Pang , Manxi Song , Jiahao Yang , Haobin Mo , Kai Wang , Xia Chen , Yujun Huang , Ruixia Gu , Chengran Guan
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Abstract

Lysozyme, an antimicrobial agent, is extensively employed in the food and healthcare sectors to facilitate the breakdown of peptidoglycan. However, the methods to improve its catalytic activity and secretory expression still need to be studied. In the present study, twelve lysozymes from different origins were heterologously expressed using the Komagataella phaffii expression system. Among them, the lysozyme from the European flat oyster Ostrea edulis (oeLYZ) showed the highest activity. Via a semi-rational approach to reduce the structural free energy, the double mutant Y15A/S39R (oeLYZdm) with the catalytic activity 1.8-fold greater than that of the wild type was generated. Subsequently, different N-terminal fusion tags were employed to enhance oeLYZdm expression. The fusion with peptide tag 6×Glu resulted in a remarkable increase in the recombinant oeLYZdm expression, from 2.81 × 103 U mL–1 to 2.11 × 104 U mL–1 in shake flask culture, and eventually reaching 2.05 × 105 U mL–1 in a 3-L fermenter. The work produced the greatest amount of heterologous oeLYZ expression in microbial systems that are known to exist. Reducing the structural free energy and employing the N-terminal fusion tags are effective strategies to improve the catalytic activity and secretory expression of lysozyme.

Abstract Image

从欧洲平牡蛎(Ostrea edulis)中高效生产高活性溶菌酶。
溶菌酶是一种抗菌剂,被广泛用于食品和医疗保健领域,以促进肽聚糖的分解。然而,提高其催化活性和分泌表达的方法仍有待研究。本研究利用 Komagataella phaffii 表达系统异源表达了 12 种不同来源的溶菌酶。其中,来自欧洲平牡蛎(Ostrea edulis)的溶菌酶(oeLYZ)活性最高。通过降低结构自由能的半理性方法,产生了催化活性比野生型高 1.8 倍的 Y15A/S39R 双突变体(oeLYZdm)。随后,利用不同的 N 端融合标签来提高 oeLYZdm 的表达。与肽标签 6×Glu 融合后,重组 oeLYZdm 的表达量显著增加,在摇瓶培养中从 2.81 × 103 U-mL-1 增加到 2.11 × 104 U-mL-1,最终在 3 升发酵罐中达到 2.05 × 105 U-mL-1。这是目前已知微生物系统中异源表达 oeLYZ 数量最多的一次。降低结构自由能和使用 N 端融合标签是提高溶菌酶催化活性和分泌表达的有效策略。
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来源期刊
Journal of biotechnology
Journal of biotechnology 工程技术-生物工程与应用微生物
CiteScore
8.90
自引率
2.40%
发文量
190
审稿时长
45 days
期刊介绍: The Journal of Biotechnology has an open access mirror journal, the Journal of Biotechnology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. The Journal provides a medium for the rapid publication of both full-length articles and short communications on novel and innovative aspects of biotechnology. The Journal will accept papers ranging from genetic or molecular biological positions to those covering biochemical, chemical or bioprocess engineering aspects as well as computer application of new software concepts, provided that in each case the material is directly relevant to biotechnological systems. Papers presenting information of a multidisciplinary nature that would not be suitable for publication in a journal devoted to a single discipline, are particularly welcome.
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