Multifunctional alkalophilic α-amylase with diverse raw seaweed degrading activities

• Published in: AMB Express 2021-10, Vol.11 (1), p.139-139, Article 139
• Main Authors: Gu, Xiaoqian, Fu, Liping, Pan, Aihong, Gui, Yuanyuan, Zhang, Qian, Li, Jiang
• Format: Article
• Language: English
• Subjects: Agarase; Algae; Alginate lyase; Alginates; Alginic acid; Amino acids; Amylases; Biomedical and Life Sciences; Biotechnology; Cellulase; Coliforms; Degradation; Digestion; Domains; E coli; Enzymatic digestion; Industrial applications; Life Sciences; Metagenomic; Metagenomics; Microbial Genetics and Genomics; Microbiology; Multifunctional enzyme; Oligosaccharides; Original; Original Article; Seaweeds; Substrates; Sugar; Thermal stability; α-Amylase
• ISSN: 2191-0855; 2191-0855
• DOI: 10.1186/s13568-021-01300-x

Uncultured microbes are an important resource for the discovery of novel enzymes. In this study, an amylase gene ( amy2587 ) that codes a protein with 587 amino acids (Amy2587) was obtained from the metagenomic library of macroalgae-associated bacteria. Recombinant Amy2587 was expressed in Escherichia coli BL21 (DE3) and was found to simultaneously possess α-amylase, agarase, carrageenase, cellulase, and alginate lyase activities. Moreover, recombinant Amy2587 showed high thermostability and alkali resistance which are important characteristics for industrial application. To investigate the multifunctional mechanism of Amy2587, three motifs (functional domains) in the Amy2587 sequence were deleted to generate three truncated Amy2587 variants. The results showed that, even though these functional domains affected the multiple substrates degrading activity of Amy2587, they did not wholly explain its multifunctional characteristics. To apply the multifunctional activity of Amy2587, three seaweed substrates ( Grateloupia filicina, Chondrus ocellatus , and Scagassum ) were digested using Amy2587. After 2 h, 6 h, and 24 h of digestion, 121.2 ± 4 µg/ml, 134.8 ± 6 µg/ml, and 70.3 ± 3.5 µg/ml of reducing sugars were released, respectively. These results show that Amy2587 directly and effectively degraded three kinds of raw seaweeds. This finding provides a theoretical basis for one-step enzymatic digestion of raw seaweeds to obtain seaweed oligosaccharides.