Characterization of an Alginate Lyase，from Abalone and Its Expression in Escherichia coli

doi:10.3969/j.issn.1673-1689.2018.06.010

Home > Archive>Volume 37, Issue 6, 2018 >632-638. DOI:10.3969/j.issn.1673-1689.2018.06.010

Characterization of an Alginate Lyase，from Abalone and Its Expression in Escherichia coli
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                        10.3969/j.issn.1673-1689.2018.06.010
                    
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Alginate oligosaccharides have good prospects because of their useful and biologically important functions. Alginate lyase can degrade alginate to alginate oligosaccharides，genetic engineering technology provides technology for efficient production of alginate lyase. Extraction of RNA from the hepatopancreas of abalone cDNA obtained by RT-PCR，cDNA as template for PCR amplification to obtain alginate lyase gene algl，connection with pET-28a（+） vector. The recombinant plasmid was transformed into E.coli BL21，induced and expressed. The new protein bands about 32 000 showed on polyacrylamide gel electrophoresis（SDS-PAGE） and the protein form of inclusion bodies. Inclusion body purification，refolding and determination of its activity 15.6 U/mL，specific activity 644.6 U/mL. The optimum temperature and pH were 35 ℃ and 8，respectively. Alyl only degradation poly mannuronate（polyM） and not degradable poly guluronic acid（polyG）. The Km and Vmax values for alginate were 1.71 mg/mL and 19.08 U/mL，respectively. Recombinase Algl poly mannuronic acid substrate specificity and cold adaptation has the potential for biotechnology and industral applications.

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ZHANG Qi, LI Yuntao, WANG Liping. Characterization of an Alginate Lyase，from Abalone and Its Expression in Escherichia coli[J]. Journal of Food Science and Biotechnology,2018,37(6):632-638.

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Online: September 27,2018
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