• Journal of Microbiology and Biotechnology
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• 제10권3호
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• pp.349-354
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• 2000

Abstract A facultative alkalophilic gram-negative Vibrio metschnikovii strain RH530, isolated from the wastewater, produced several alkaline proteases (VAP) including six alkaline serine proteases and a metalloprotease. From this strain, high yielding YAP mutants were isolated by NTG treatment. The isolated mutant KS1 showed nine times more activity than the wild-type after optimization of the culture media. The production was regulated by catabolite repression when glucose was added to the medium. The effects of several organic nitrogen sources on the production of the YAP were investigated to avoid catabolite repression. The combination of 4% wheat gluten meal (WGM), 1.5% cotton seed flour (eSF), and 5% soybean meal (SBM) resulted in the best production when supplemented with 1% NaCl. The YAP showed a resistance to surfactants such as $sodium-{\alpha}-olefin$ sulfonate (AOS), polyoxy ethylene oxide (POE), and sodium dodecyl sulfate (SDS), yet not to linear alkylbenzene sulfonate (LAS). However, the activity of the YAP was restored completely when incubated with LAS in the presence of POE or $Na_2SO_4$. The YAP was stable in a liquid laundry detergent containing 6.6% SLES (sodium lauryl ether sulfate), 6.6% LAS, 19.8% POE, and stabilizing agents for more than two weeks at $40^{\circ}C$, but the stability was sharply decreased even after 1 day when incubated at $60^{\circ}C$. A washing performance test with the YAP exhibited it to be a good washing power by showing 51 % and 60% activity at $25^{\circ}C{\;}and{\;}40^{\circ}C$, respectively, thereby indicating that the YAP also has a good detergency at a low temperature. All the results suggest that the YAP produced from the mutant strain KSI has suitable properties for use in laundry detergents.rgents.

• 한국균학회지
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• 제25권2호통권81호
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• pp.91-100
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• 1997

회분배양과 유가배양을 이용하여 호알칼리성 Cephalosporium sp. RYM-202로부터 alkaline carboxymethyl cellulase (CMCase)와 Xylanase의 생산을 위한 배양조건에 대하여 조사하였다. 조사한 탄소기질 중에서 밀기울이 두 효소의 생산에 가장 효율적이었다. 또한 CMCase는 carboxymethyl cellulose (CMC)를 탄소기질로 첨가한 배양액에서, 반면에 xylanase의 경우에는 xylan을 기질로 하였을 때 높은 생산량을 나타냄으로서 유도기질 특이성을 보였는 바, 이 결과는 Cephalosporium sp. RYM-202에서의 CMCaee와 xylanase의 생합성이 효소 유도 수준에서 독립적으로 조절됨을 시사해 준다. 조사된 질소원 중에서는 무기질소원인 $NaNO_3$가 효소생산에 효과적이었으며, 이들 효소의 최대 생산을 위한 배양온도와 pH는 각각 $20^{\circ}C$와 9.0인 것으로 나타났다. 한편, 발효조에서의 회분배양을 통한 효소생산의 경우, 밀기울의 농도를 5%까지 증가시킴에 따라 효소생산량은 증가되었으나 catabolite repression에 의해 효소생산의 지연과 생산력의 감소를 초래하였다. 이러한 문제점은 탄소원의 간헐적 공급에 의한 유가배양을 통해서 어느 정도 해결될 수 있는 것으로 나타났으며, 밀기울의 최종농도가 5% 되게 공급된 유가배양 시 CMCase와 xylanase의 최대 효소생산량은 각각 0.39 및 9.2 units/ml 이었으며, 이는 같은 농도의 밀기울을 함유하는 회분배양 시 획득된 효소활성에 비해 각각 1.22배와 1.36배 증가된 것이다.

• 한국균학회지
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• 제26권1호통권84호
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• pp.127-133
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• 1998

Trichoderma reesei QM 9414를 N-methyl-N'-nitro-N-nitrosoguanidine으로 처리하여 얻은 돌연변이주들을 여러 탄소원에서 배양했을 때 carboxymethylcellulose(CMC), fiter paper 등의 기질에 대한 활성측정결과 다음과 같은 결과를 얻었다. 모균주가 catabolite repression을 받는 반면에 돌연변이주들의 cellulase 활성은 glucose를 탄소원으로 사용한 배지에서 모균주에 비해 CMCase 8.4배, FPase가 $5.4{\sim}5.7$배 증가함으로써 glucose-derepression성질을 갖는 돌연변이주들을 얻었으며 glucose를 탄소원으로 사용하고 lactose로 효소의 생산을 유도시켜본 결과 모균주에 비해 효소의 생산에 안정성을 갖는 것으로 나타났으며 돌연변이주 1이 상대적으로 더 우수한 돌연변이주로 나타났다. 돌연변이주들에 의해 생산된 효소는 모균주와 마찬가지로 pH 4.8, $60^{\circ}C$에서 최적활성을 나타내었다.

• Journal of Microbiology and Biotechnology
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• 제6권2호
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• pp.103-109
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• 1996

Mn(II)+succinate decreased the carotenoid formation of the yeast Phaffia rhodozyma, probably by scavenging $O_2$. When duroquinone (DQ), an internal and external $O_2$ generator, was added to medium, P. rhodozyma produced more amount of carotenoids. The increased carotenoid production was destroyed by oxygen radical (OR) scavengers, ascorbate+Cu(II) and dimethylsulfoxide. When sub-lethal concentrations of $H_2O_2$ , an external OR source, and antimycin, an internal OR inducer, were used, the effect of $H_2O_2$ on carotenoid formation and composition was less significant than that of antimycin. Addition of superoxide dismutase, an external OR remover, rescued cells from death caused by the high concentration of DO. In this condition, the yeast culture showed an increase in carotenoid content. Addition of DQ into P. rhodozyma culture in the stationary phase did not increase carotenoid production. Therefore, carotenoid formation was stimulated by internal ORs in the growing yeast. It was probably due to release of catabolite repression on carotenogenesis in the yeast. Aeration was important for carotenoid production but was not as effective as the internal OR producer, DQ.

• Journal of Microbiology and Biotechnology
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• 제17권12호
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• pp.2081-2084
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• 2007

A 2.5 kb aga gene encoding ${\alpha}$-galactosidase (${\alpha}$-Gal) from Leuconostoc mesenteroides SY1 was cloned into pSJE, an E. coli-Leuconostoc shuttle vector. The recombinant plasmid, pSJEaga, was introduced into Leuconostoc citreum KCTC3526 (ATCC49370) by electroporation. Transcription level of aga was the highest in cells grown on raffinose (1%, w/v) followed by cells grown on galactose, melibiose, fructose, glucose, and sucrose. Western blot using antibodies against ${\alpha}$-Gal showed similar results to slot-blot results and enzyme activity measurements. All the results indicated that the aga was successfully expressed in L. citreum and its transcription was under the carbon catabolite repression (CCR).

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.259-262
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• 2000

The production of pullulan by Aureobasidium pullulans HP-2001 was investigated under various ratios of glucose as carbon source and yeast extract as the nitrogen source, Highest conversion rate (productivity) of glucose to pullulan was 40.0% when concentrations of glucose and yeast extract were 5% and 0.15%, respectively. Maximal production of pullulan was 29.3g/1 when the concentration of glucose was 8%(w/v) and that of yeast extract was 40:1. On basis of the result that production of pullulan was found in a medium which concentration of glucose as carbon source was up to 20%(w/v), Aureobasidium pullulans HP-2001 seemed to overcome the catabolite repression. Conversion rate of pullulan from 20%(w/v) of glucose was 11.1%.

• Journal of Microbiology and Biotechnology
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• 제6권1호
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• pp.12-18
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• 1996

A chitosanase-producing bacteria was isolated on chitosan agar plate from soil samples. The strain was spore-forming gram positive bacteria, catalase positive, and rod shape. The strain was identified as Bacillus cereus. The strain did not need an inducer for the synthesis of chitosanase. Chitosanase from Bacillus sp. HW-002 was constitutive enzyme. The optimal medium for the production of the enzyme was composed of 0.5$\%$ sucrose and $1.5\%$ yeast extract-tryptone (1:1 w/w) mixture at pH 6.5. After Bacillus sp. HW-002 was cultivated at $32^{\circ}C$ for 32 h, maximal productivity was gained to be about 27, 200 U/l. Chitosanase from Bacillus sp. HW-002 was a mixed growth-linked metabolite.

• 한국미생물·생명공학회지
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• 제34권4호
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• pp.289-298
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• 2006

3',5'-cyclic adenosine monophosphate (cAMP) is an important molecule, which mediates diverse cellular processes. For example, it is involved in regulation of sugar uptake/catabolism, DNA replication, cell division, and motility in various acterial species. In addition, cAMP is one of the critical regulators for syntheses of virulence factors in many pathogenic bacteria. It is believed that cAMP acts as a signal for environmental changes as well as a regulatory factor for gene expressions. Therefore, intracellular concentration of cAMP is finely modulated by according to its rates of synthesis (by adenylate cyclase), excretion, and degradation (by cAMP phosphodiesterase). In the present review, we discuss the bacterial physiological characteristics governed by CAMP and the molecular mechanisms for gene regulation by cAMP. Furthermore, the effect of cAMP on phosphotransferase system is addressed.

• KSBB Journal
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• 제13권2호
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• pp.203-208
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• 1998

We investigated the effects of dissolved oxygen (D.O.) and fructose (C-source) on cell growth and biosynthesis of cyclosporin A (CyA) produced as a secondary metabolite by a wild-type filamentous fungus, Tolypocladium inflatum. This was performed by controlling the level of D.O. and the residual C-source, as required, through adjustment of medium flow rate, medium concentration and agitation rate in fed-batch cultures. CyA production was furned out to be maximal, when D.O. level was controlled around 10% saturated D.O. and concentration of the C-source was maintained sufficiently low (below 2 g/L) not to cause carbon catabolite repression. Under this culture condition, we obtained the highest values of CyA concentration (507.14 mg/L), Qp (2.11 mg CyA/L/hr), $Y_x/s$ (0.49 g DCW/g fructose), $Y_p/s$<(22.56 mg CyA/g fructose), and YTEX>$_p/x$ (48.31 mg CyA/g DCW), but relatively lower values of cell concentration (11.98 g DCW/L) and cell productivity (0.043 g DCW/L/hr), in comparison with other parallel fed-batch fermentation conditions. These results implied that, in the carbon-limited culture with 10% saturated D.O. level, the producer microorganism utilized the C-source more efficiently for secondary metabolism.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.746-753
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• 2008

The hprK gene encoding bifunctional HPrK/P (kinase/phosphorylase) was cloned from L. mesenteroides SY1, a strain isolated from kimchi. hprK was transcribed as a monocistronic gene. His-tagged HPrH16A and HPrK/P were produced in E. coli BL21 (DE3) using pET26b(+) and purified. HPrK/P phosphorylation assay with purified proteins showed that the kinase activity of HPrK/P increased at slightly acidic pHs. Divalent cations such as $Mg^{2+}$ and $Mn^{2+}$ and glycolytic intermediates such as fructose-1, 6-bisphosphate (FBP) and phosphoenolpyruvate (PEP) increased the kinase activity of HPrK/P, but inorganic phosphate strongly inhibited it. Kinetic studies for the kinase activity of HPrK/P showed that the apparent $K_m$ values were 0.18 and $14.57{\mu}M$ for ATP and HPr, respectively. The $K_m$ value for the phosphorylase activity of HPrK/P was $14.16{\mu}M$ for P-Ser-HPr (HPr phosphorylated at the serine residue).