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Thermostability enhancement of chitosanase CsnA by fusion a family 5 carbohydrate-binding module

Author:
Han, Yujuan, Gao, Peixin, Yu, Wengong, Lu, Xinzhi
Source:
Biotechnology letters 2017 v.39 no.12 pp. 1895-1901
ISSN:
0141-5492
Subject:
carbohydrate binding, catalytic activity, chitosanase, protein secondary structure, thermal stability
Abstract:
OBJECTIVE: To determine the effects of carbohydrate-binding modules (CBMs) on the thermostability and catalytic efficiency of chitosanase CsnA. RESULTS: Three CBMs (BgCBM5, PfCBM32-2 and AoCBM35) were engineered at the C-terminus of chitosanase CsnA to create hybrid enzymes CsnA-CBM5, CsnA-CBM32 and CsnA-CBM35. K ₘ values of all the hybrid enzymes were lower than that of the wild type (WT) enzyme; however, only CsnA-CBM5 had an elevated specific activity and catalytic efficiency. The fusion of BgCBM5 enhanced the thermostability of the enzyme, which exhibited a 8.9 °C higher T₅₀ and a 2.9 °C higher Tₘ than the WT. Secondary structural analysis indicated that appending BgCBM5 at the C-terminus considerably changed the secondary structure content. CONCLUSIONS: The fusion of BgCBM5 improved the thermal stability of CsnA, and the obtained hybrid enzyme (CsnA-CBM5) is a useful candidate for industrial application.
Agid:
5852897