Identification of a novel alkaline amylopullulanase from a gut metagenome of Hermetia illucens
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- 作者：Young-Seok Lee ; So-Hyeon Seo ; Sang-Hong Yoon ; Su-Yeon Kim ; Bum-Soo Hahn ; Joon-Soo Sim ; Bon-Sung Koo ; Chang-Muk Lee ; changmuk@rda.go.kr" class="auth_mail" title="E-mail the corresponding author ; changmuk@hotmail.com" class="auth_mail" title="E-mail the corresponding author
- 关键词：Pullulanase ; Hermetia illucens ; Metagenomic library ; Purification ; Characterization
- 刊名：International Journal of Biological Macromolecules
- 出版年：2016
- 出版时间：January 2016
- 年：2016
- 卷：82
- 期：Complete
- 页码：514-521
- 全文大小：1383 K
文摘
A novel pullulanase gene, PulSS4, was identified from the gut microflora of Hermetia illucens by a function-based metagenome screening. The PulSS4 gene had an open reading frame of 4455 base pairs, and encoded a mature protein of 1484 amino acids, with a signal peptide sequence of 44 amino acids. The deduced amino acid sequence of PulSS4 gene showed 51% identity with that of the amylopullulanase of Amphibacillus xylanus, exhibiting no significant sequence homology to already known pullulanases. A conserved domain analysis revealed it to be a pullulanase type II with respective active sites at the N-terminal pullulanase and C-terminal amylase domain. PulSS4 was active in the temperature range of 10–50 °C, with an optimum activity at 40 °C. It was active in the pH range of 6.5–10.5, with optimum pH at 9.0, and retained more than 80% of its original activity in a broad pH range of 5–11 for 24 h at 30 °C. Also, PulSS4 was highly stable against many different chemical reagents, including 10% polar organic solvents and 1% non-ionic detergents. Overall, PulSS4 is expected to have the strong potential for application in biotechnological industries that require high activity at moderate temperature and alkaline conditions.