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来自[具体来源未给出]的5家族纤维素酶在BL21-CodonPlus(DE3)-RIPL中的重组表达及特性分析

Recombinant expression and characterization of the family 5 cellulase from in BL21-CodonPlus (DE3)-RIPL.

作者信息

Tran Dinh Minh, Nguyen Thi Huyen, Huynh To Uyen, Pentekhina Iuliia

机构信息

Institute of Biotechnology and Environment, Tay Nguyen University, Buon Ma Thuot, Dak Lak, 630000, Viet Nam.

Institute of Biotechnology, Bioengineering and Food Systems, Advanced Engineering School, Far Eastern Federal University, Vladivostok, 690922, Russia.

出版信息

Biochem Biophys Rep. 2024 Dec 14;41:101898. doi: 10.1016/j.bbrep.2024.101898. eCollection 2025 Mar.

Abstract

B. velezensis RB. IBE29 is a chitinolytic bacterium originally isolated from agricultural soil of Vietnam. Previous studies demonstrated this bacterium was a promising chitinase producer, biocontrol agent, and biofertilizer for crop production. The complete genome sequence of the bacterium was reported and possesses the gene encoding family 5 cellulase; however, role of this enzyme has not been experimentally characterized. This work aimed to express and biologically characterize family 5 cellulase of strain RB. IBE29. The ORF (without signal peptide) of the of strain RB. IBE29 was expressed in BL21-CodonPlus (DE3)-RIPL using the vector pColdII, and the corresponding product (rBvCelA, 55.17 kDa) was successfully purified using the HisTrap FF column. The purified rBvCelA showed the highest cellulase activity against CMC, followed by sugarcane bagasse and rice straw, and had optimal temperature and pH at 60 °C and 6.0, respectively. Metal salts ZnCl, FeCl, MgCl, CuSO, and MnCl enhanced the cellulase activity by 103.85, 124.24, 109.38, 105.64, and 115.12 %, respectively. In addition, the supplementation of the purified rBvCelA in the feed enhanced the growth and improved the feed intake of broiler chickens by 5.88 and 5.19 %, respectively. These results indicated that family 5 cellulase of has a promising role in crop production and poultry breeding. As far as we know, this is the first report describing the contribution of family 5 cellulase from in broiler breeding.

摘要

贝莱斯芽孢杆菌RB. IBE29是一种最初从越南农业土壤中分离出来的几丁质分解细菌。先前的研究表明,这种细菌是一种有前景的几丁质酶生产者、生物防治剂和用于作物生产的生物肥料。该细菌的完整基因组序列已被报道,并且拥有编码5类纤维素酶的基因;然而,这种酶的作用尚未通过实验进行表征。这项工作旨在表达并对RB. IBE29菌株的5类纤维素酶进行生物学表征。RB. IBE29菌株的(无信号肽)开放阅读框使用载体pColdII在BL21-CodonPlus(DE3)-RIPL中表达,并且使用HisTrap FF柱成功纯化了相应产物(rBvCelA,55.17 kDa)。纯化后的rBvCelA对羧甲基纤维素表现出最高的纤维素酶活性,其次是甘蔗渣和稻草,其最佳温度和pH分别为60℃和6.0。金属盐ZnCl、FeCl、MgCl、CuSO和MnCl分别使纤维素酶活性提高了103.85%、124.24%、109.38%、105.64%和115.12%。此外,在饲料中添加纯化后的rBvCelA分别使肉鸡的生长速度提高了5.88%,采食量提高了5.19%。这些结果表明,RB. IBE29的5类纤维素酶在作物生产和家禽养殖中具有广阔的应用前景。据我们所知,这是第一份描述RB. IBE29的5类纤维素酶在肉鸡养殖中的作用的报告。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad3c/11699454/9b7ca142f4cb/gr1.jpg

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