Expression and Characterization of a Novel Cold-Adapted and Stable β-Agarase Gene agaW1540 from the Deep-Sea Bacterium Shewanella sp. WPAGA9

• Published in: Marine drugs Vol. 19; no. 8; p. 431
• Main Authors: Wang, Wenxin, Wang, Jianxin, Yan, Ruihua, Zeng, Runying, Zuo, Yaqiang, Wang, Dingquan, Qu, Wu
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 29.07.2021; MDPI
• Subjects: Adaptation; Agarase; Agarose; Amino acids; Animals; Aquatic Organisms - chemistry; Bacteria; Bacterial Proteins - genetics; Biodegradation; Biological activity; Chromatography; Cold; Cold Temperature; cold-adapted; Copper; Deep sea; Deep sea environments; Deep water; Degradation; Enzymatic activity; Enzyme activity; Genomes; Hydrogen-Ion Concentration; Incubation period; Industrial production; Iron; Molecular weight; neoagaro-oligosaccharides; Oligosaccharides; pH effects; Phylogenetics; Recombinants; Room temperature; Sepharose - genetics; Shewanella; Shewanella - genetics; stable; Temperature; Thin layer chromatography; Working conditions; Zinc; β-agarase; Beijing China; China; neoagaro-oligosaccharides; deep-sea; β-agarase; cold-adapted; stable
• ISSN: 1660-3397; 1660-3397
• DOI: 10.3390/md19080431

The neoagaro-oligosaccharides, degraded from agarose by agarases, are important natural substances with many bioactivities. In this study, a novel agarase gene, agaW1540, from the genome of a deep-sea bacterium sp. WPAGA9, was expressed, and the recombinant AgaW1540 (rAgaW1540) displayed the maximum activity under the optimal pH and temperature of 7.0 and 35 °C, respectively. rAgaW1540 retained 85.4% of its maximum activity at 0 °C and retained more than 92% of its maximum activity at the temperature range of 20-40 °C and the pH range of 4.0-9.0, respectively, indicating its extensive working temperature and pH values. The activity of rAgaW1540 was dramatically suppressed by Cu and Zn , whereas Fe displayed an intensification of enzymatic activity. The and of rAgaW1540 for agarose degradation were 15.7 mg/mL and 23.4 U/mg, respectively. rAgaW1540 retained 94.7%, 97.9%, and 42.4% of its maximum activity after incubation at 20 °C, 25 °C, and 30 °C for 60 min, respectively. Thin-layer chromatography and ion chromatography analyses verified that rAgaW1540 is an endo-acting β-agarase that degrades agarose into neoagarotetraose and neoagarohexaose as the main products. The wide variety of working conditions and stable activity at room temperatures make rAgaW1540an appropriate bio-tool for further industrial production of neoagaro-oligosaccharides.