• Journal of Microbiology and Biotechnology
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• v.15 no.1
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• pp.59-65
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• 2005

Poly-${\gamma}$-glutamic acid (PGA) is an unusual anionic polypeptide and has great potential as an environmentally and industrially significant biodegradable material. A new ${\gamma}$-PGA producer, Bacillus mesentericus MJM1, with high production capacity was isolated from Korean domestic Chungkuckjang bean paste. It produced ${\gamma}$-PGA at the level of 10 g/l in suitable media. The viscosities of 5% initially extracted mucin and purified ${\gamma}$-PGA solutions were 660 cps and 600 cps, respectively. The produced ${\gamma}$-PGA polymer consisted of 2,000 glutamic acid residues with even proportion of L and D types with molecular mass of about 200- 300 kDa. Bacillus mesentericus MJM1 displayed ${\gamma}$-glutamyltranspeptidase (${\gamma}$-GTP) activity that is known to play a key role in ${\gamma}$-PGA biosynthesis. The ${\gamma}$-GTP coding region was located on the plasmid of 5.8 kb. The plasmid, named pMMH1, is a rolling-circle replication (RCR) plasmid and additionally contained a replication origin and type I signal peptidase (sipP) coding region.

• Journal of Microbiology
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• v.40 no.2
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• pp.140-145
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• 2002

The 5.8 kb pMMH1, rolling-circle replication (RCR) plasmid of the wild type soil Bacillus mesentericus was developed into a novel secretion vector system in Bacillus subtilis. The pMMHl turned out to have a replication origin and two open reading frames (ORFs) of the putative γ-GTP and type I signal peptidase (sipP). To characterize the regions necessary for plasmid stability and high copy number, five vectors (pPS, pPP, pEN, pMN, pME) were constructed by disruption or deletion of each region in pMMH1. Like pMMHl all constructed vectors were stable over 100 generations In a non-selective medium. Since pPS was the smallest (2.3 kb)of all, it was selected for the construction of a navel secretion vector, Using the $\alpha$-amylase promoter/signal sequence of B. subtilils the novel plasmid pJSN was constructed. When $\beta$-glucosidase was expressed using pJSN, we found $\beta$-glucosidase activity in the medium. This result strongly suggested that plasmid pJSN can be used for the production of bioactive peptides in B. subtilis.

• Journal of Aquaculture
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• v.11 no.4
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• pp.495-503
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• 1998

The present study investigated the effects of Bisroot, that contains live bacteria (Bacillus polyfermenticus, Bacillus mesentericus, Streptococcus faecalis, & Bifidobacterium breve) and digestive enzymes (protease, lipase), on the growth, body composition and immune response of Nile tilapia fingerlings. One percent of the Bisroot was added to the experimenta feed. All exprimental fish were fed for 60 days. The weigh gains among the experimental fish were not significntly different (P>0.05). Hematocrit value, hemoglobin, total protein, glucose, GOT, and GPT were unaffected by Bisroot treatment. However, it was observed that glucose, GOT, and GPT value in the fish that were fed Bisroot, were lower than the control. The complement activity ($CH_50$) tended to be significantly increased by Bisroot treatment, but not lysozyme activity. Phagocytosis and respiratory burst activities of macrophages in the head kidney were enhanced by Bisroot. Therefore, the Bisroot diet enhances the cellular immune activities were enhanced by Bisroot. Therefore, the Bisroot diet enhances the cellular immune activities of non-specific immune responses. When fish were challenged with a virulent strain of Edwardsiella tarda, the Bisroot treated fish were more resistant than the control. The present results suggest that the introduction of Bisroot into the diet of Nile tilapia could increase their resistance against bacterial infection, reduce fish mortality, and offers economic benefits.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.348-354
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• 2004

The expression and antibacterial. activity of recombinant human lactoferrin (hLf) was studied from meth­ylotrophic yeast Pichia pastoris. The gene encoding hLf, isolated from human breast cDNA library, was subcloned into the expression vector, pPIC3.5K under the control of AOX1 promoter. The gene was integrated into the host chromosome and was identified by Southern blotting. The expression of the integrated gene was investigated by RT-PCR, Northern blotting, SDS-PAGE and Western blotting. Discrete band corresponding to hLf was detected from the SDS-PAGE, which was confirmed by Western blotting. The expression was also confirmed by RT-PCR and Northern blotting. The antibacterial activity of the recombinant hLf (rhLf) was investigated using Staphy­lococcus aureus ATCC 6538P and Micrococcus flavus ATCC 10240 as test organisms. The rhLf showed strong antibacterial activities against the bacteria. Furthermore, many Gram-negative animal pathogens such as E.coli ATCC8739, 25922, and Salmonella typhimurium 114 and 115, Pseudomonas fluorescens ID 963 I, P. aeruginosa KCCM 11802, and Gram-positive bacteria Bacillus mesentericus were also inhibited in their growth by the rhLf.