College of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, People's Republic of China.
Prep Biochem Biotechnol. 2022;52(3):273-282. doi: 10.1080/10826068.2021.1934698. Epub 2021 Jun 10.
The chaperone-assisted soluble expression and characterization of high molecular weight chitinase chiZJ408 in BL21 were investigated. Chitin degradation products by chitinase chiZJ408 were analyzed. The results indicated that the introduction of the chaperone GroELS promoted the correct folding of chitinase chiZJ408 and increased its soluble expression by 14.9% ( < 0.05) in BL21. The optimal pH and temperature of chitinase chiZJ408 were respectively 6.0 and 50 °C. Chitinase chiZJ408 was stable at pHs of 4.0 ∼ 7.0 and at below 40 °C. Mgand Ca had a significant impact on improving the activity of chitinase chiZJ408. Chitinase chiZJ408 was demonstrated to be exochitinase that cleaved the -1,4-glycosidic bond of the chitin chain to generate only -diacetylchitobiose. This study broadens our understanding of chitinase and provides a basis for solving the problem of inclusion body formed by long fragment gene expression in .
研究了在 BL21 中辅助伴侣蛋白可溶性表达和高相对分子质量几丁质酶 chiZJ408 的特性。分析了几丁质酶 chiZJ408 对几丁质降解产物的作用。结果表明，伴侣蛋白 GroELS 的引入促进了几丁质酶 chiZJ408 的正确折叠，使其在 BL21 中的可溶性表达增加了 14.9%（<0.05）。几丁质酶 chiZJ408 的最适 pH 和温度分别为 6.0 和 50°C。几丁质酶 chiZJ408 在 pH4.0∼7.0 和低于 40°C 时稳定。Mg 和 Ca 对提高几丁质酶 chiZJ408 的活性有显著影响。几丁质酶 chiZJ408 被证明是一种外切几丁质酶，它能切断几丁质链的-1,4-糖苷键，只生成二乙酰基壳二糖。本研究拓宽了我们对几丁质酶的认识，为解决长片段基因表达形成包涵体的问题提供了依据。
