A Novel Agarase, Gaa16B, Isolated from the Marine Bacterium Gilvimarinus agarilyticus JEA5, and the Moisturizing Effect of Its Partial Hydrolysis Products

• Published in: Marine drugs 2021-12, Vol.20 (1), p.2
• Main Authors: Lee, Youngdeuk, Jo, Eunyoung, Lee, Yeon-Ju, Eom, Tae-Yang, Gang, Yehui, Kang, Yoon-Hyeok, Marasinghe, Svini Dileepa, Hettiarachchi, Sachithra Amarin, Kang, Do-Hyung, Oh, Chulhong
• Format: Article
• Language: English
• Subjects: Agarase; Agarose; Aging; Amino acids; Animals; Aquatic Organisms; Bacteria; Carbohydrates; Chromatography; Cloning; Cosmeceuticals; Cosmetics; E coli; Gammaproteobacteria; Gels; Genomes; Gilvimarinus agarilyticus JEA5; Glycosidases; Glycoside hydrolase; glycoside hydrolase family 16; Glycoside Hydrolases - chemistry; Glycoside Hydrolases - pharmacology; Glycosides; Hyaluronic acid; hyaluronidase inhibition; Hydrogen-Ion Concentration; Hydrolase; Hydrolysis; Molecular weight; neoagaro-oligosaccharide; Oligosaccharides; partial hydrolytic product; pH effects; Polymers; Proteins; Recombinants; Skin; Thin layer chromatography; β-agarase
• ISSN: 1660-3397; 1660-3397
• DOI: 10.3390/md20010002

We recently identified a β-agarase, Gaa16B, in the marine bacterium JEA5. Gaa16B, belonging to the glycoside hydrolase 16 family of β-agarases, shows less than 70.9% amino acid similarity with previously characterized agarases. Recombinant Gaa16B lacking the carbohydrate-binding region (rGaa16Bc) was overexpressed in and purified. Activity assays revealed the optimal temperature and pH of rGaa16Bc to be 55 ∘C and pH 6-7, respectively, and the protein was highly stable at 55 ∘C for 90 min. Additionally, rGaa16Bc activity was strongly enhanced (2.3-fold) in the presence of 2.5 mM MnCl . The and of rGaa16Bc for agarose were 6.4 mg/mL and 953 U/mg, respectively. Thin-layer chromatography analysis revealed that rGaa16Bc can hydrolyze agarose into neoagarotetraose and neoagarobiose. Partial hydrolysis products (PHPs) of rGaa16Bc had an average molecular weight of 88-102 kDa and exhibited > 60% hyaluronidase inhibition activity at a concentration of 1 mg/mL, whereas the completely hydrolyzed product (CHP) showed no hyaluronidase at the same concentration. The biochemical properties of Gaa16B suggest that it could be useful for producing functional neoagaro-oligosaccharides. Additionally, the PHP of rGaa16Bc may be useful in promoting its utilization, which is limited due to the gel strength of agar.