Isolation of Novel Exo-type β-Agarase from Gilvimarinus chinensis and High-level Secretory Production in Corynebacterium glutamicum

Abstract

Agar, a major cell wall component in marine red macroalgae, has recently gained high interest as a potential renewable biomass. By the activity of β-agarase which cleaves the β-1,4-glycosidic bond, agarose can be hydrolyzed into neoagarobioses which have great potential in the cosmetics, food, and medical industries. Here, based on sequence homology analysis, we isolated a novel exotype β-agarase (EXB3) from Gilvimarinus chinensis which have putative glycoside hydrolase (GH) 50 domain. The optimum pH and temperature for the activity of EXB3 were pH 7.0 and 30°C, respectively. The Km and Vmax for agarose were 26 mg/mL and 126.8 U/mg, respectively, and the Kcat/Km value was 3.7 × 105 s-1M-1. Under the optimal condition (30°C and pH 7), it was clearly confirmed that neoagarobiose (NA2) was produced as a major product directly from agarose. For the large-scale production of EXB3, we also developed a secretory production platform in Corynebacterium glutamicum. During the fed-batch cultivation in 2 L-scale bioreactor, EXB3 was successfully produced in the culture medium as high as 458.3 mg/L, and EXB3 was purified from the culture supernatant with high purity and recovery yield (24%).

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Correspondence to Ki Jun Jeong.

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Jeong, Y.J., Choi, J.W., Cho, M.S. et al. Isolation of Novel Exo-type β-Agarase from Gilvimarinus chinensis and High-level Secretory Production in Corynebacterium glutamicum. Biotechnol Bioproc E 24, 250–257 (2019). https://doi.org/10.1007/s12257-018-0362-x

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Keywords

  • β-agarase
  • Gilvimarinus chinensis
  • Corynebacterium glutamicum
  • secretion
  • fed-batch cultivation