Zeng Rong, Yin Xiao-Yan, Ruan Tao, Hu Qiao, Hou Ya-Li, Zuo Zhen-Yu, Huang Hao, Yang Zhong-Hua
College of Chemistry and Chemical Engineering, Hubei University, Wuhan 430062, China.
College of Chemical Engineering and Technology, Wuhan University of Science and Technology, Wuhan 430081, China.
Bioengineering (Basel). 2016 Apr 19;3(2):13. doi: 10.3390/bioengineering3020013.
Screening and obtaining a novel high activity cellulase and its producing microbe strain is the most important and essential way to improve the utilization of crop straw. In this paper, we devoted our efforts to isolating a novel microbe strain which could produce high activity cellulase. A novel strain ZY-01 was isolated from a cropland where straw is rich and decomposed, by using the soil dilution plate method with cellulose and Congo red. The strain has been licensed with a patent numbered ZL 201210295819.6. The cellulase activity in the cultivation broth could reach up to 7.4 IU/mL at a non-optimized fermentation condition with the newly isolated ZY-01. The cellulase was separated and purified from the culture broth through (NH₄)₂SO₄ fractional precipitation, anion-exchange chromatography and gel filtration chromatography. With the separation process, the CMC specific activity increased from 0.88 IU/mg to 31.5 IU/mg with 35.8 purification fold and 47.04% yield. Furthermore, the enzymatic properties of the cellulase were investigated. The optimum temperature and pH is 50 °C and pH 5.0 and it has good thermal stability. Zn, Ca and Mn could remarkably promote the enzyme activity. Conversely, Cu and Co could inhibit the enzymatic activity. This work provides a new highly efficient strain for cellulase production and shows good prospects in practical application.
筛选并获得一种新型高活性纤维素酶及其产生菌是提高作物秸秆利用率的最重要且必不可少的途径。本文致力于分离一种能够产生高活性纤维素酶的新型菌株。通过使用纤维素和刚果红的土壤稀释平板法，从富含秸秆且已分解的农田中分离出了一种新型菌株ZY - 01。该菌株已获得专利授权，专利号为ZL 201210295819.6。在使用新分离的ZY - 01进行非优化发酵条件下，发酵液中的纤维素酶活性可达7.4 IU/mL。通过硫酸铵分级沉淀、阴离子交换色谱和凝胶过滤色谱从发酵液中分离纯化纤维素酶。在分离过程中，CMC比活性从0.88 IU/mg提高到31.5 IU/mg，纯化倍数为35.8，产率为47.04%。此外，还研究了纤维素酶的酶学性质。其最适温度和pH分别为50℃和pH 5.0，并且具有良好的热稳定性。锌、钙和锰能显著促进酶活性。相反，铜和钴会抑制酶活性。这项工作为纤维素酶生产提供了一种新的高效菌株，并在实际应用中显示出良好的前景。
