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通过改良固态发酵系统生产单宁酶的新型优化策略

Novel optimization strategy for tannase production through a modified solid-state fermentation system.

作者信息

Wu Changzheng, Zhang Feng, Li Lijun, Jiang Zhedong, Ni Hui, Xiao Anfeng

机构信息

1College of Food and Biological Engineering, Jimei University, Xiamen, 361021 China.

Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen, 361021 Fujian China.

出版信息

Biotechnol Biofuels. 2018 Apr 2;11:92. doi: 10.1186/s13068-018-1093-0. eCollection 2018.

Abstract

BACKGROUND

High amounts of insoluble substrates exist in the traditional solid-state fermentation (SSF) system. The presence of these substrates complicates the determination of microbial biomass. Thus, enzyme activity is used as the sole index for the optimization of the traditional SSF system, and the relationship between microbial growth and enzyme synthesis is always ignored. This study was conducted to address this deficiency. All soluble nutrients from tea stalk were extracted using water. The aqueous extract was then mixed with polyurethane sponge to establish a modified SSF system, which was then used to conduct tannase production. With this system, biomass, enzyme activity, and enzyme productivity could be measured rationally and accurately. Thus, the association between biomass and enzyme activity could be easily identified, and the shortcomings of traditional SSF could be addressed.

RESULTS

Different carbon and nitrogen sources exerted different effects on microbial growth and enzyme production. Single-factor experiments showed that glucose and yeast extract greatly improved microbial biomass accumulation and that tannin and (NH)SO efficiently promoted enzyme productivity. Then, these four factors were optimized through response surface methodology. Tannase activity reached 19.22 U/gds when the added amounts of tannin, glucose, (NH)SO, and yeast extract were 7.49, 8.11, 9.26, and 2.25%, respectively. Tannase activity under the optimized process conditions was 6.36 times higher than that under the initial process conditions. The optimized parameters were directly applied to the traditional tea stalk SSF system. Tannase activity reached 245 U/gds, which is 2.9 times higher than our previously reported value.

CONCLUSIONS

In this study, a modified SSF system was established to address the shortcomings of the traditional SSF system. Analysis revealed that enzymatic activity and microbial biomass are closely related, and different carbon and nitrogen sources have different effects on microbial growth and enzyme production. The maximal tannase activity was obtained under the optimal combination of nutrient sources that enhances cell growth and tannase accumulation. Moreover, tannase production through the traditional tea stalk SSF was markedly improved when the optimized parameters were applied. This work provides an innovative approach to bioproduction research through SSF.

摘要

背景

传统固态发酵(SSF)系统中存在大量不溶性底物。这些底物的存在使微生物生物量的测定变得复杂。因此,酶活性被用作优化传统SSF系统的唯一指标,而微生物生长与酶合成之间的关系一直被忽视。本研究旨在解决这一不足。用水提取茶梗中的所有可溶性营养物质。然后将水提取物与聚氨酯海绵混合,建立改良的SSF系统,用于生产单宁酶。利用该系统,可以合理、准确地测定生物量、酶活性和酶生产率。因此,可以很容易地确定生物量与酶活性之间的关联,并解决传统SSF的缺点。

结果

不同的碳源和氮源对微生物生长和酶产生有不同的影响。单因素实验表明,葡萄糖和酵母提取物极大地促进了微生物生物量的积累,单宁和硫酸铵有效地提高了酶生产率。然后,通过响应面法对这四个因素进行了优化。当单宁、葡萄糖、硫酸铵和酵母提取物的添加量分别为7.49%、8.11%、9.26%和2.25%时,单宁酶活性达到19.22 U/gds。优化工艺条件下的单宁酶活性比初始工艺条件下高6.36倍。优化后的参数直接应用于传统茶梗SSF系统。单宁酶活性达到245 U/gds,比我们之前报道的值高2.9倍。

结论

本研究建立了一种改良的SSF系统,以解决传统SSF系统的缺点。分析表明,酶活性与微生物生物量密切相关,不同的碳源和氮源对微生物生长和酶产生有不同的影响。在促进细胞生长和单宁酶积累的营养源最佳组合下,获得了最大的单宁酶活性。此外,应用优化参数后,传统茶梗SSF的单宁酶产量显著提高。这项工作为通过SSF进行生物生产研究提供了一种创新方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a129/5879601/a3ab9f8488fa/13068_2018_1093_Fig1_HTML.jpg

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