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Home > Archive>Volume 36, Issue 9, 2017 >938-943. DOI:10.3969/j.issn.1673-1689.2017.09.008
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Effects of Different Types of Peptide Linkers on the Enzymatic Properties of β-Mannanase
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    Abstract:

    A family 27 carbohydrate-binding module（CBM27） of Thermotoga maritima MSB8 was linked to the C-terminus of a glycoside hydrolase family 5 β-mannanaseof Aspergillus usamii YL-01-78（AuMan5A） by using a flexible peptide linker F（GGGGS）3 ora rigid peptide linker R（EAAAK）3. By overlapping PCR technique，twofusion enzyme genes，Auman5A-F-cbm27 and Auman5A-R-cbm27，was amplified. The gene Auman5A and two fusion genes were expressed in Pichia pastoris GS115，respectively. And then，the enzymatic properties of the recombinant β-mannanases，reAuMan5A，reAuMan5A-F-C and reAuMan5A-R-C，were analyzed. As a result，the Km ofreAuMan5A-R-C towards locust bean was 0.9 mg/mL，which was much smaller than those of reAuMan5A（1.7 mg/mL） and reAuMan5A-F-C（1.9 mg/mL）. The temperature optimum（Topt） of reAuMan5A-F-C was 65 ℃，which was 5 ℃ lower than that（70 ℃） of reAuMan5A or reAuMan5A-R-C. The half-lives at 70℃（t701/2） of reAuMan5A-F-C and reAuMan5A-R-C were 36 and 124 min，respectively，which were about 4- and 13.8 folds longer than that（t701/2=9 min） of reAuMan5A. In this study，the fusionenzymereAuMan5A-R-C was obtained with strong substrate affinity，high thermostability and wide range of pH stability. It will be a potential candidate for industrial processes，such as food，animal feed，energy，pharmaceuticals and so on.

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WANG Chunjuan, TANG Shihan, WU Minchen, DONG Yunhai, HANGYiling, LI Jianfang. Effects of Different Types of Peptide Linkers on the Enzymatic Properties of β-Mannanase[J]. Journal of Food Science and Biotechnology,2017,36(9):938-943.

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