• Korean Journal of Microbiology
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• v.42 no.2
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• pp.142-148
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• 2006

In order to develop chitosanase for the production of chitosan oligosaccharides, a chitosanase-producing bacterium was isolated from the traditional fermented soybean, Meju, and identified as Bacillus amyloliquefaciene MJ-1. The cloned chitosanase gene, 825 bp in size, encoded a single peptide of 274 amino acids with a estimated molecular mass of 30.9 kDa. The deduced amino acid sequence showed significant homology with microbial chitosanases. The recombinant chitosanase was expressed in Escherichia coli upon induction with isopropyl-D-thiogalactopyranoside, and purified using $Ni^{2+}-NTA$ agarose column chromatography. The maximal activity of the recombinant chitosanase is at pH 5.0 and $60^{\circ}C$. The recombinant chitosanase is stable between pH 5.0 and pH 7.0 at $37^{\circ}C$ for 30 min, and more than 75% of the activity still remain at $80^{\circ}C$ for 30 min incubation.

• The Korean Journal of Food And Nutrition
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• v.24 no.1
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• pp.124-131
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• 2011

We analyzed content and peptides in order to investigate the productivity from Cheonggookjang(fast-fermented soybean paste), fermenting it for 180 hours at $40^{\circ}C$. Results showed that pH was 7.07 at the start and became 7.41 in 24 hours, it eventually increased to 8.63 after 180 hours. Acidity was 0.2 in 12 hours, 0.5 in 12 hours, and then remained on 0.1 thereafter. Total sugar was 1.54 mg/$m\ell$ at the start, but it gradually decreased to 0.76 mg/$m\ell$ after the lapse of 48 hours, and 1.0 mg/$m\ell$ in 120 hours, and finally 0.8 mg/$m\ell$ in 180 hours. Reducing sugar was 0.14 mg/$m\ell$ at the start, and 0.88 mg/$m\ell$ after the lapse of 24 hours, 0.64 mg/$m\ell$ in 48 hours, 0.26 mg/$m\ell$ in 72 hours, and showed no definite change untill 180 hours. The amount of free amino acid was $0.19\;{\mu}M/\ell$ at the start, and $4.88\;{\mu}M/\ell$ after the lapse of 72 hours, $4.5\;{\mu}M/\ell$ in 120 hours, and then it rapidly decreased to $0.23\;{\mu}M/\ell$ after180 hours. Absorbance of soluble protein and peptide at 280 nm was 12.4 in 48 hours, 31.12 in 120 hours, and 31.12 in 180 hours. HPLC revealed that in the fermentation process, large molecular proteins are hydrolyzed into small peptides and amino acids, and after the lapse of 48 hours the pattern became almost the same. The protease activity of Cheonggookjang was 0.011 unit/$m\ell$ after the lapse of 36 hours and then it decreased. The result shows as Cheonggookjang started its deamination of amino acid in 100 hours, it is desirable to produce peptide within 100 hours of its fermentation.