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中国精品科技期刊2020
吴诗丽,谢莹莹,段雪凝,等. 产纤维素酶菌株的筛选及其酶学性质研究[J]. 食品工业科技,xxxx,x(x):1−9. doi: 10.13386/j.issn1002-0306.2024080165.
引用本文: 吴诗丽,谢莹莹,段雪凝,等. 产纤维素酶菌株的筛选及其酶学性质研究[J]. 食品工业科技,xxxx,x(x):1−9. doi: 10.13386/j.issn1002-0306.2024080165.
WU Shili, XIE Yingying, DUAN Xuening, et al. Screening of Cellulase Producing Strains and Study on Their Enzymatic Properties[J]. Science and Technology of Food Industry, xxxx, x(x): 1−9. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2024080165.
Citation: WU Shili, XIE Yingying, DUAN Xuening, et al. Screening of Cellulase Producing Strains and Study on Their Enzymatic Properties[J]. Science and Technology of Food Industry, xxxx, x(x): 1−9. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2024080165.

产纤维素酶菌株的筛选及其酶学性质研究

Screening of Cellulase Producing Strains and Study on Their Enzymatic Properties

  • 摘要: 为获得高效降解纤维素的菌种及酶资源,以传统发酵食品(白酒糟和酱醪)源微生物为研究对象。采用刚果红染色法平板筛选和3,5-二硝基水杨酸法酶活筛选产纤维素酶菌株,并对菌株进行菌落形态特征观察、生理生化特征试验及分子生物学鉴定,探究菌株在不同胁迫条件下产酶情况,以及菌株所产纤维素酶的酶学性质。结果表明,共筛选出了3株纤维素酶活相对较高的菌株,经鉴定均为贝莱斯芽孢杆菌(Bacillus velezensis),其羧甲基纤维素酶活力为109.37±6.89~125.97±4.37 U/mL。不同胁迫条件下菌株产酶性能表明:随着盐含量的增加,菌株酶活力呈下降趋势,当盐含量大于9%,酶活力仍能保留在20 U/mL,且随着盐含量的进一步提高,酶活力基本保持不变;在0~8%的乙醇含量范围内,菌株酶活力呈先上升后下降的趋势,3株菌的酶活均大于80 U/mL,其中,菌株B12在乙醇含量为6%时酶活力最高,菌株JL39、JL55在乙醇含量4%时酶活力最高。纤维素酶酶学性质研究表明:菌株胞外纤维素酶最适反应温度为50 ℃;最适pH为5.5~6.5;金属离子Cu2+、Ca2+、Mg2+对3株菌的酶活力都有促进作用,其中Cu2+的促进作用最强,而Fe2+、Mn2+、Fe3+和Ni+对酶活力的作用各有所异。本研究为产纤维素酶菌株资源的开发利用提供了菌种资源,为抗逆性纤维素酶的高效应用提供了宝贵的酶资源,为生物质资源的转化和利用奠定了基础。

     

    Abstract: In order to obtain efficient cellulose degrading strains and enzyme resources, the microorganisms from traditional fermented food (white wine lees and sauce mash) were used as the research objects. The cellulase-producing strains were screened by plate screening using Congo red staining method and enzyme activity screening using 3,5-dinitrosalicylic acid method, and the cellulase-producing strains were subjected to colony morphological observation, physiological and biochemical characteristics and molecular biology identification, to investigate the enzyme production of the strains under different stress conditions, as well as the enzymatic properties of the cellulase produced by the strains. The results showed that a total of three strains with relatively high cellulase activity were screened, all of which were identified as Bacillus velezensis, with carboxymethylcellulase activity ranging from 109.37±6.89 U/mL to 125.97±4.37 U/mL. The enzyme production performance of the strains under different stress conditions showed that the enzyme activity of the strains showed a decreasing trend with the increase of salt content. When the salt content was greater than 9%, the enzyme activity could still be retained at 20 U/mL, and with the further increase of salt content, the enzyme activity basically remained unchanged; in the range of 0~8% ethanol content, the enzyme activity of the strains showed an increasing and then decreasing trend, and the enzyme activity of all three strains were greater than 80 U/mL, among which, strain B12 had the highest enzyme activity at 6% ethanol content, and strains JL39, JL55 had the highest enzyme activity at 4% ethanol content. The enzymatic properties of cellulase showed that: the optimal reaction temperature of extracellular cellulase was 50 ℃; the optimal pH was 5.5~6.5; the metal ions Cu2+, Ca2+, and Mg2+ had a promotional effect on the enzyme activity of three strains, of which Cu2+ had the strongest promotional effect, while Fe2+, Mn2+, Fe3+, and Ni+ had a different effect on the enzyme activity. This study provides strain resources for the development and utilization of cellulase-producing strain resources, provides valuable enzyme resources for the efficient application of resistant cellulases, and lays the foundation for the transformation and utilization of biomass resources.

     

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