• Microbiology and Biotechnology Letters
• /
• v.26 no.1
• /
• pp.34-39
• /
• 1998

We have already prepared a human lysozyme (HLY) structural gene from chemically synthesized 38 oligomers with high codon usage in Saccharomyces cerevisiae. For directing the synthesis and secretion of HLY in S. cerevisiae, two types of expression vectors, a YCp centromere-based vector, pHK101 and a YEp 2-$\mu\textrm{m}$ circle-based vector, pHK501 were constructed. With the resulting plasmids, we have confirmed that yeast transformant harboring pHK501 has more secreted HLY than pHK101-transformant by using a lysoplate and a turbidimetric assay. In flask cultivation, pHK501-transformant produced active HLY about 8 times (55 units/$m\ell$) higher than pHK101-transformant. From batch cultivation, the HLY productivity was obtained with 1.12 units/$m\ell$/h, corresponding to a 1.8-fold increase compared with flask fermentation. These results indicate that yeast transformant with pHK501 vector overexpressed and secreted HLY than that of YCp type vector.

• Korean Journal of Food Science and Technology
• /
• v.23 no.2
• /
• pp.167-174
• /
• 1991

Cultural conditions for the biopolymer production by an alkali tolerant Bacillus sp. isolated from soil were investigated and determination of optimal conditions was carried out by response surface method. The maximal production of biopolymer was obtained after cultivation at $30^{\circ}C$ for 36hrs in the mixture of 8% soluble starch, 0.75% yeast extract, 0.1% $NaNO_3$, 0.05% $MgSO_4\;7H_2O$ and 1% $Na_2CO_3$ adjusted to pH 10. Under these conditions, about 44 g/l of biopolymer were produced. From the results of response surface analysis, optimal condition for the production of biopolymer were obtained at stationary point with 15.16 of C/N ratio, $34.62^{\circ}C$ of temperature and 9.50 of pH. On the basis of these conditions, it was estimated that 66.84 g/l of the biopolymer could be produced.

• Applied Biological Chemistry
• /
• v.40 no.6
• /
• pp.561-566
• /
• 1997

Screening, Identification and some cultural characteristics of ALA$({\delta}-aminolevulinic\;acid)$-producing photosynthetic bacteria were carried out for the optimal production of ALA, one of the bioherbicides. Among photosynthetic bacteria isolated from soil, marsh, pond, etc., KK-10 was the best producer of ALA and identified to be Rhodobacter capsulatus belonging to a typical group of nonsulfur purple bacteria. By addition of 15 mM LA (levulinic acid), an inhibitor of ALA dehydrase in cyclic tetrapyrrole biosynthesis, into culture broth at middle log phase of cell growths, ALA production was considerably increased to about 20-fold (28 mg/l). The combined supplementation of glycine and succinate, each with a concentration of 30 mM also enhanced production of ALA and activity of ALA synthase to about 50-fold (73 mg/l) and 2-fold, respectively. The isolated strain was able to produce upto 80 mg/l under the cultural condition optimized by addition 15 mM LA into the synthetic medium at four different points starting middle log phase.

• Microbiology and Biotechnology Letters
• /
• v.20 no.4
• /
• pp.477-482
• /
• 1992

A bacterial strain Acinetobacter calcoaceticus was examined for its ability to degrade fats, oils and hydrocarbons, and tested for the possibility of application in wastewater treatment. All fats and oils tested were degraded by the strain. About 60% of hexadecane, 26% of fish oiL and 40-54% of vegetable oils were consumed respectively in shaking-flask culture. Saturated fatty acid compositions were about 55% in fish oil and 6-12% in vegetable oils. Increases in cell mass were accompanied with decreases in the concentrations of carbon sources. When jar fermentor in place of shaking-flask was used as a culturing vessel. above 80% of all carbon sources was consumed and yield of cell mass was improved to nearly 1.00. Synthetic wastewaters containing 3% of fat, oil, or hydrocarbon as a sale ca,bon source were treated sequentially with A. calcoaceticus first and then exposed to activated sludge. The concentrations of carbon sources were decreased below 0.06% through the process, and the concentrations of suspended solids were lower than 53 mglml. The data imply the potential use of A. calcoaceticus in wastewater treatment.

• Korean Journal of Food Science and Technology
• /
• v.26 no.5
• /
• pp.490-495
• /
• 1994

Enzymatic solubilization of tofu-residue was attempted using carbohydrase isolated from Aspergillus niger CF-34. Tofu-residue, by-product of tofu manufacture or soymilk processing was used as the model for plant cell wall. It was found that tofu-residue was rich in nurients: 46.7% carbohydrate, 32.8% protein, the rest being lipid and ashes. Carbohydrate component of tofu-residue consisted of 36.8% cellulose and 62.6% hemicellulose. The carbohydrase was found to consist of pectinase, xylanase, PGase, CMCase, and SFase when tofu-residue and pectin were used as the carbon source. Enzyme induction was maximum at 7days of culture. Optimum reaction pH was 4.0, temperature $50^{\circ}C$. The enzyme was stable to $50^{\circ}C$, above which the stability decreased rapidly.

• Microbiology and Biotechnology Letters
• /
• v.20 no.3
• /
• pp.335-343
• /
• 1992

A Streptomyces strain No. 182-27, which produced amino acid antimetabolite, was isolated from soil. During the course of screening for new amino acid antimetabolites from the culture broths of Actinomycetes, we found that the strain produced a substance active against Gram-positive bacteria and its activity was reversed by L-Ieucine on the synthetic minimal agar medium in the culture broth. The morphological and cultural characteristics serve to identify the producing organism strain 182-27 as the Streptomyces, although the species of this strain should be resolved in further studies. Fermentation was carried out in the synthetic medium at $28^{\circ}C$ for 78 hours. The fermentation yield reached about 2 mg per liter of the broth. Purification was done by ion exchange resin, active carbon, silica gel column chromatography and obtained 20 mg of pure active substance from the 20 $\ell$ culture broth. The 182-27 substance was obtained as white powder, mp 18SoC. From the physicochemical characteristics of the substance, it was amino acid like substance but unknown about its chemical structure. It is active against some Gram-positive bacteria and reversed by L-Ieucine.

• Korean Journal of Food Science and Technology
• /
• v.31 no.6
• /
• pp.1628-1634
• /
• 1999

The polymerase chain reaction (PCR) for the sensitive and specific detection of Listeria monocytogenes was employed by using LM 1 and LM 2 primers which were based on the listeriolysin O gene. The direct use of cell suspension as DNA template, without DNA extraction or lysis step, was suitable and specific enough to detect L. monocytogenes at the level of $10^2$ CFU or less per PCR for the pure culture and milk sample, however, the detection sensitivity became blunt for other food samples such as kimchi and chicken. The nested PCR, in which L-1 and L-2 (both designed from listeriolysin O gene) were employed as inner primers, was specific for detecting L. monocytogenes and enhanced the detection limit by 10 times. The PCR using LM 1 and LM 2 primers was very effective to detect L. monocytogenes from foods in terms of the specificity and time consumed, i. e. within $4{\sim}8\;hrs$ (nested PCR).

• KSBB Journal
• /
• v.15 no.5
• /
• pp.428-433
• /
• 2000

Cell cultures of Camptotheca acuminata, which is known to produce the anticancer indole alkaloid camptothecin and its dervatives, were made to enhance the productivity of camptothecin. In suspension cultures, the maximum cell growth rate in exponential growth phase was $0.269day^{-1}$ which was correlated to 2.58days of cell doubling time. The production of camptothecin was non-growth associated. The camptothecin production was the highest at 11th day form inoculation and then it decreased. Various elicitors were applied to enhance the production of camptothecin. Both of jasmonic acid and cellulase increased the production of camptothecin. The optimal dosing time of elicitor was the beginning of the cultures. The combination of two elicitors was more effective to produce camptothecin than single applications. $20.42{\times}10^{-3}mg/\ell$ of camptothecin was obtained with combined application of two elicitors in two days.

• Proceedings of the Korean Aquaculture Society Conference
• /
• 2003.10a
• /
• pp.147-148
• /
• 2003

동해안의 강릉 연안에 자생하는 토종 다시마인 개다시마는 산업적으로 개발의 여지가 매우 크다. 그러나 천연 자원량에 의존하고 있는 가공산업은 해황에 따라 원료 공급이 좌우되는 불안정성을 가지고 있어서 이 식물의 산업적 생산량을 확보하기 위한 방안으로 대량양식의 필요성이 대두되고 있다. 본 연구는 동해안 토종 다시마인 개다시마의 종묘생산기술 개발을 위해서 항온배양기와 실내수조배양을 실시하고 바다 가이식 양성에 의한 환경적응성 실험을 수행하였다. 연구 방법은 2001년 10월에 강릉연안 수심 20m에서 분포하는 개다시마 군락의 모조로부터 유주자액을 받아 온도 5, 10, 15, 20, $25^{\circ}C$, 조도 40, 80, 120 $\mu$mol m$^{-2}$ s$^{-1}$의 조건에서 6주간 정치배양하였다. 유주자의 발아 및 아포체 생장 실험 이후 바다 가이식 및 양성실험은 2002년 1~2월까지 1개월간 실시하였다. 유주자의 발아 및 아포체 형성은 1$0^{\circ}C$, 120$\mu$mo1 m$^{-2}$ s$^{-1}$에서 배양 2주만에 가장 먼저 아포체의 발아가 관찰되었으며, 이후 15$^{\circ}C$와 5$^{\circ}C$에서 순차적으로 아포체의 발아가 이루어졌다. 2$0^{\circ}C$는 80, 120$\mu$mol m$^{-2}$ s$^{-1}$에서 소수의 아포체만이 관찰되었으며, $25^{\circ}C$에서는 아포체가 형성되지 않았다. 아포체는 1$0^{\circ}C$에서 가장 좋은 생장을 보였고, 15$^{\circ}C$와 1$0^{\circ}C$에서는 비슷한 생장을 나타냈으며, 2$0^{\circ}C$에서는 80$\mu$mol m$^{-2}$ s$^{-1}$에서만 아포체의 형성이 관찰되어 가장 낮은 생장을 보였다. 동일 온도에서는 조도에 따른 생장의 차이가 많이 났는데, 40$\mu$mol m$^{-2}$ s$^{-1}$에서 가장 좋은 생장을 나타냈다. 80과 120$\mu$mol m$^{-2}$ s$^{-1}$에서는 120$\mu$mol m$^{-2}$ s$^{-1}$이 약간 좋은 생장을 보였지만 큰 차이는 나지 않았다. 생장이 가장 좋은 조건은 1$0^{\circ}C$, 40$\mu$mol m$^{-2}$ s$^{-1}$으로 엽장 1.28mm까지 생장했다. 전체적으로 수심이 깊은 곳에 서식하는 특성상 저수온과 저조도에서 좋은 생장 경향을 보였다. 실내배양을 통해 얻은 엽장 약 1~2mm의 종묘를 2002년 1월말에 삼척 호산에 위치한 바다 양성장에서 종묘들과 함께 2주간 수심별로 가이식(수온 12$^{\circ}C$)을 실시한 결과, 수심 1m에서 좋게 나타나 엽장이 약 5mm정도 생장하였다. 수심 0m에서는 강한 조도에 의해 엽상체가 사그라드는 기형이 나타났으며, 수심 5m에서는 엽상체의 기형이나 생장의 차이는 없었지만, 색택이 약간 바랜 것을 볼 수 있었다. 종합적으로 볼 때 종묘의 관리나 생장에는 수심 1~2m가 양호할 것으로 생각된다. 바다 가이식을 통해 엽장 5mm의 종묘로 생장시킨 후 바로 본양성장에 씨줄감기를 하여 양성을 실시하였다 양성 1개월 후 엽상체는 약 2~3cm, 최대 4~5cm까지 생장하였다. 이를 통해 강릉연안 토종 다시마인 개다시마에 대한 종묘 생산과 대량 양성의 가능성을 확인하였다.

• Journal of Life Science
• /
• v.19 no.10
• /
• pp.1478-1483
• /
• 2009

A solid-state culture based on grain materials was attempted to produce a fibrinolytic enzyme for blood circulation improvement using Bacillus subtilis BK-17. The spore, rather than vegetative cell inoculation, of B. subtilis BK-17 on the solid-state culture was effective in the production of a fibrinolytic enzyme. Maximum spore production was obtained with a SFM medium (0.8% nutrient broth, 0.05% yeast extract, $10^{-1}$ M $MgCl_2$, $10^{-3}$ M $FeCl_3$, $10^{-4}$ $MnCl_2$, $10^{-5}$ M dipicolic acid, pH 6.5). Optimal pH and temperature were pH 6 and $30^{\circ}C$, respectively. The spore production reached a maximum at 60 hours of incubation. Bacillus subtilis BK-17 on the mung bean solid-state culture produced greater fibrinolytic activity, and less activity was seen in other grains such as kidney bean, soybean and corn. Protein and lipid contents of fermented soybeans were about 10 - 30% more than those of unfermented soybeans. Amino acid content was also 5 - 20% more than that of unfermented soybeans. These results indicated that fermented solid-state culture medium, fermented soybean in this case, can be utilized as a food supplement.