• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.91-94
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• 2002

Arachidonic acid is a polyunsaturated fatty acid(PUFA) containing twenty carbon atoms with four double bonds. The family of w-6 PUFA, including arachidonic acid as well as r-linoleic acid, was served as intermediates in the formation of several key prostaglandin and leukotrienes. Several fungal strains of the genus Mortierella accumulate high amounts of arachidonic acid. In this study experiments were carried out to optimize the culture conditions for the mass production of fungus Mortierella alpina DSA -12 and lipid production with high proportion of polyunsaturated fatty acids, especially arachidonic acid. The batch culture was carried out in 500 L fermenter containing 50 g/L glucose, 18 g/L corn-steep powder and 100 mg/L MnS04 under $25^{\circ}C$, aeration rate of 0.5 vvm and agitation speed of 200 rpm without pH control. As a result, we could be obtained 22 g/L of cell mass with high contents of lipid 12.1 g/L) and arachidonic acid (5.1 g/L) The intermittent fed-batch culture was performed in the medium containing 20 g/L glucose and 10 g/L corn-steep powder. The final glucose concentration was 170 g/L and pH was maintained at 5.5 ${\sim}$ 6.0 by adding 14% ammonia solution. It was shown relatively high cell concentration (70.5 g/L) with high contents of lipid (45.8 g/L) and arachidonic acid 08.3 g/L). Therefore, when compared to batch cultures, the high concentration of arachidonic acid could be obtained by fed-batch culture using M. alpina DSA -12. These results imply that the fed-batch culture of M. alpina DSA -12 was feasible in industrial purpose and could be employed in the commercial production of arachidonic acid.

• 한국토양비료학회지
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• 제49권5호
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• pp.461-468
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• 2016

One of the important phytohormones produced by plant growth promoting bacteria is the auxin; indole-3-acetic acid (IAA), with L-tryptophan as the precursor. In this study, we focused on the investigation of optimal conditions for the production of IAA by Bacillus megaterium BM5. We investigated culturing conditions, such as incubation temperature, pH of the culture medium and incubation period, with varying media components such as inoculation volume, tryptophan concentration and carbon and nitrogen source. Besides, optimization study intended for high IAA production was carried out with fermentation parameters such as rpm and aeration. The initial yield of $42{\mu}g\;IAA\;ml^{-1}$ after 24 hr increased to $85{\mu}g\;ml^{-1}$ when 5% (v/v) of L-tryptophan was used in the culture broth. The maximum yield of $320{\mu}g\;IAA\;ml^{-1}$ was observed in trypticase soy broth (TSB) supplemented with starch and soybean meal as C and N sources with a C/N ratio of 3:1 (v/v) at $30^{\circ}C$, pH 8.0 for 48 hrs with 1.0 vvm and 250 rpm in 5 L working volume using 10 L scale fermenter. The bacterial auxin extracted from the culture broth was confirmed by thin layer chromatography and high-performance liquid chromatography and effect on plant growth was confirmed by root elongation test.

• 한국미생물·생명공학회지
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• 제43권2호
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• pp.164-168
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• 2015

효모의 액침배양법에 의한 구연산을 발효생산하였다. 여덟 가지 구연산 발효균주의 생산성을 비교하여 가장 생산성이 높은 Candida zeylanoides를 선발하였다. 발효조건의 최적화를 위하여 glucose와 fructose 혼합당 배지에서 C/N 비, 인산염의 농도, Fe²⁺ 이온의 최적 농도를 결정하였다. 삼각 플라스크 실험결과 C/N 비는 40/1이 가장 좋았으며, 최적화된 인삼염과 Fe²⁺ 농도에서 구연산 91.4 g/l와 이소-구연산 19.8 g/l를 생성하였다. 이 결과를 토대로 TCPH 전처리 당밀 배지를 사용한 실험에서는 인삼염 농도 1.0 g/l, Fe²⁺ 이온 50 mg/l 이하의 농도에서 구연산 91 g/l와 이소-구연산 18.5 g/l를 생성하였다. 5 L-Jar 발효조를 사용한 실험에서의 구연산 수율(Y_p/s, g/g)은 0.51이었다.

• Journal of Microbiology and Biotechnology
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• 제23권2호
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• pp.189-194
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• 2013

In a study of hydrogen-producing bacteria, strain T4384 was isolated from rice field samples in the Republic of Korea. The isolate was identified as Enterobacter sp. T4384 by phylogenetic analysis of 16S rRNA and rpoB gene sequences. Enterobacter sp. T4384 grew at a temperature range of $10-45^{\circ}C$ and at an initial pH range of 4.5-9.5. Strain T4384 produced hydrogen at 0-6% NaCl by using glucose, fructose, and mannose. In serum bottle cultures using a complete medium, Enterobacter sp. T4384 produced 1,098 ml/l $H_2$, 4.0 g/l ethanol, and 1.0 g/l acetic acid. In a pH-regulated jar fermenter culture with the biogas removed, 2,202 ml/l $H_2$, 6.2 g/l ethanol, and 1.0 g/l acetic acid were produced, and the lag-phase time was 4.8 h. Strain T4384 metabolized the hydrolysate of organic waste for the production of hydrogen and volatile fatty acid. The strain T4384 produced 947 ml/l $H_2$, 3.2 g/l ethanol, and 0.2 g/l acetic acid from 6% (w/v) food waste hydrolysate; 738 ml/l $H_2$, 4.2 g/l ethanol, and 0.8 g/l acetic acid from Miscanthus sinensis hydrolysate; and 805 ml/l $H_2$, 5.0 g/l ethanol, and 0.7 g/l acetic acid from Sorghum bicolor hydrolysate.

• 한국초지조사료학회지
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• 제36권4호
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• pp.344-349
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• 2016

본 연구에서는 주로 한(육)우, 낙농 등에서 이용되는 총체맥류 조사료를 돼지, 닭 등에게까지 확대 이용 가능성을 모색하기 위하여 수행하였다. 이를 위해 총체맥류를 분쇄할 수 있는 시스템을 개발하였고 분쇄시스템을 통해 분쇄한 총체맥류를 비닐 백에 밀봉 저장한 후 사료가치를 분석하였다. 총체맥류는 Fig. 4에서 보는 바와 같이 수확 직후 줄기, 잎, 곡실 등을 분리하지 않고 원형 그대로 분쇄장치에 투입하였다. 분쇄되어 배출된 총체맥류의 분쇄상태를 분석한 결과 알곡 20%, 10 mm 이상 4%, 5~10 mm 27%, 5 mm 이하 49%로 매우 미세하게 분쇄되는 것으로 나타나 단위동물의 배합사료에 일정비율 혼합하여 급이 하는 것이 가능할 것으로 판단되었다. 분쇄 직후 밀봉하여 상온에서 저장한 총체맥류 사일리지는 약 3개월이 지난 시점에서 품질을 분석하였다. 시험에 사용한 총체맥류는 새쌀보리와 금강밀이다. 분쇄된 총체맥류의 발효를 촉진하기 위하여 하나에는 발효제를 접종하였고 다른 하나는 발효제를 접종하지 않았다. 발효제를 접종한 분쇄 총체 새쌀보리의 조단백질, 조지방, 조섬유, NDF, ADF, 리그닌, 셀룰로스 함량은 2.45%, 1.61%, 8.95%, 16.94%, 9.52%, 1.01%, 8.51%로 나타났으며 TDN, RFV는 81.38%, 447.5%로 나타났다. 발효제를 접종하지 않은 총체 새쌀보리의 조단백질, 조지방, 조섬유, NDF, ADF, 리그닌, 셀룰로스 함량은 2.57%, 1.62%, 9.61%, 18.25%, 10.13%, 1.10%, 9.04%로 나타났으며 TDN, RFV는 80.90%, 412.9%로 나타났다. 발효제를 접종한 분쇄 총체 금강밀의 조단백질, 조지방, 조섬유, NDF, ADF, 리그닌, 셀룰로스 함량은 2.43%, 1.27%, 10.99%, 19.49%, 11.23%, 1.46%, 9.77%로 나타났으며 TDN, RFV는 80.03%, 382.6%로 나타났다. 발효제를 접종하지 않은 총체금강밀의 조단백질, 조지방, 조섬유, NDF, ADF, 리그닌, 셀룰로스 함량은 2.28%, 1.44%, 10.08%, 18.02%, 10.44%, 1.26%, 9.18%로 나타났으며 TDN, RFV는 80.65%, 416.9%로 나타났다. 발효제를 접종한 것과 발효제를 접종하지 않은 분쇄 총체맥류 사일리지의 상대적 사료가치를 평가한 결과 새쌀보리의 경우 발효제를 첨가한 것이 높았으며, 금강밀의 경우 발효제를 첨가하지 않은 것이 높은 것으로 나타났다. 따라서 총체맥류 분쇄 사일리지를 조제할 때 품종에 따라 발효제의 첨가 여부를 결정하는 것이 중요할 것으로 판단되었다. 또한 Ju 등(2009, 2011)의 TDN과 RFV를 비교 분석한 결과 발효제의 접종 여부와 관계없이 분쇄 총체맥류 사일리지의 TDN과 RFV가 월등히 높은 것으로 나타났다. 따라서 총체맥류 사일리지 조제시 수확${\rightarrow}$분쇄${\rightarrow}$밀봉의 과정을 거쳐 사일리지를 조제하는 것이 조사료 이용효율을 높일 수 있을 것으로 판단된다.

• 한국근적외분광분석학회:학술대회논문집
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• 한국근적외분광분석학회 2001년도 NIR-2001
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• pp.3104-3104
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• 2001

The research described here was undertaken with the aim of monitoring, optimizing and ultimately controlling the production of heterofermentative microbes used as starters in the salami industry. The use of starter cultures in the fermented meats industry is a well-established technique used to shorten and standardize the ripening process, and to improve and control the organoleptic quality of the final product. Starter cultures are obtained by the submerged cultivation of suitable microorganisms in stirred, and sometimes aerated, fermenters where monitoring of key physiological parameters such as the concentration of biomass, substrates and metabolites suffers from the general lack of real-time measurement techniques applicable to aseptic processes. In this respect, the results of the present work are relevant to all submerged fermentation processes. Previous work on the application of on-line NIR spectroscopy to the lactic acid fermentation (Dosi et al. - Monreal NIR1995) had successfully used a system based on a measuring cell included in a circulation loop external to the fermenter. The fluid handling and sterility problems inherent in an external circulation system prompted us to explore the use of an in-line system where the NIR probe is immersed in the culture and is thus exposed to the hydrodynamic conditions of the stirred and aerated fluid. Aeration was expected to be a potential source of problems in view of the possible interference of air bubbles with the measurement device. The experimental set-up was based on an in-situ sterilizable NIR probe connected to the instrument by means of an optical fiber bundle. Preliminary work was carried out to identify and control potential interferences with the measurement, in particular the varying hydrodynamic conditions prevailing at the probe tip. We were successful in defining the operating conditions of the fermenter and the geometrical parameters of the probe (flow path, positioning, etc.) were the NIR readings were reliable and reproducible. The system thus defined was then used to construct and validate calibration curves for tile concentration of biomass, carbon source and major metabolites of two different microorganisms used as salami starters. Real-time measurement of such parameters coupled with the direct interfacing of the NIR instrument with the PC-based measurement and control system of the fermenter enabled the development of automated strategies for the interactive optimization of the starter production process.

• 한국식품과학회지
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• 제30권2호
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• pp.425-433
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• 1998

Bifidobacteria의 고농도 배양을 위한 배양액내 완충제로써 $CaCO_3$ 비드를 효율적으로 이용하기 위하여 유기산에 대한 비드의 반응특성과 유기산과 반응한 비드의 재사용 방안을 수학적 모델을 적용하여 검토하였다. 각각 다른 크기의 $CaCO_3$ 비드를 0.1 M의 혼합 유기산 용액과 Bifidobacterium longum ATCC 15707이 접종된 배양액에 각각 첨가하여 반응시킨 후, 비드 표면으로부터 감소한 $CaCO_3$ 양을 수학적 모델에 의하여 산출한 비드의 직경값과 micrometer를 이용하여 반응한 비드의 직경을 직접 측정한 값을 비교한 결과 서로 일치하였다. 그러므로, 수학적 모델이 유기산과 반응하는 $CaCO_3$ 비드의 반응특성을 설명하는데 유용하게 이용되어 질 수 있음을 확인하였다. 각각 다른 크기의 $CaCO_3$ 비드를 완충제로 사용한 배양액에 Bifidobacterium longum을 접종하여 $37^{\circ}C$로 20시간 배양한 후, $CaCO_3$ 비드의 완충효과와 직경의 변화 및 Bifidobacterium longum에 대한 증식도를 측정한 결과 비드의 직경이 작을수록 더높은 완충효과와 균증식도를 나타내었으며, 또한 비드의 직경이 크게 감소하였다. 균배양액으로부터 회수한 비드의 직경을 조사하여 반응된 비드의 전체표면적을 산출한 후 초기 사용한 비드의 전체표면적과 같도록 새로운 비드를 첨가하여 bifidobacteria를 배양한 결과 초기 사용한 비드의 완충효과와 유사한 결과를 나타내었으며, 또한 배양액내 균증식도도 같은 수준으로 증식되었음을 알 수 있었다. 따라서 bifidobacteria 배양에 완충제로 이용된 비드를 균배양에 완충제로 재차 이용할 경우 비드가 일정한 크기를 갖고, 감소한 표면적과 같은 양의 새로운 비드를 첨가한다면 사용된 비드의 재활용 가능성도 있다고 사료되었다.

• Journal of Microbiology and Biotechnology
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• 제27권3호
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• pp.500-506
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• 2017

To investigate the potential applications of bacterial heme, aminolevulinic acid synthase (HemA) was expressed in a Corynebacterium glutamicum HA strain that had been adaptively evolved against oxidative stress. The red pigment from the constructed strain was extracted and it exhibited the typical heme absorbance at 408 nm from the spectrum. To investigate the potential of this strain as an iron additive for swine, a prototype feed additive was manufactured in pilot scale by culturing the strain in a 5 ton fermenter followed by spray-drying the biomass with flour as an excipient (biomass: flour = 1:10 (w/w)). The 10% prototype additive along with regular feed was supplied to a pig, resulting in a 1.1 kg greater increase in weight gain with no diarrhea in 3 weeks as compared with that in a control pig that was fed an additive containing only flour. To verify if C. glutamicum-synthesized heme is a potential electron carrier, lactic acid bacteria were cultured under aerobic conditions with the extracted heme. The biomasses of the aerobically grown Lactococcus lactis, Lactobacillus rhamosus, and Lactobacillus casei were 97%, 15%, and 4% greater, respectively, than those under fermentative growth conditions. As a potential preservative, cultures of the four strains of lactic acid bacteria were stored at $4^{\circ}C$ with the extracted heme and living lactic acid bacterial cells were counted. There were more L. lactis and L. plantarum live cells when stored with heme, whereas L. rhamosus and L. casei showed no significant differences in live-cell numbers. The potential uses of the heme from C. glutamicum are further discussed.

• Journal of Microbiology and Biotechnology
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• 제8권5호
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• pp.536-539
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• 1998

The anti-complementary activity (immuno-stimulating activity) was tested for the exo-polymers (extra-cellular polymer) produced from submerged mycelial cultures of 21 types of higher fungi. Anti-complementary activity of the exo-polymer from Cordyceps militaris showed the highest (69.0%) followed by Pleurotus ostreatus (63.7%) and Trametes suaveolens (61.4%). The mycelial growth rate and biomass doubling time of C. militaris in a 5 I air-lift fermenter were 0.0255 $h^{-1}$ and 27.2 h, respectively. The yield of the exo-polymer produced from the culture broth of C. militaris was 2.95 mg of dry weight/ml of culture broth. Sugar and amino acid compositions of this exo-polymer were analyzed.

• Journal of Microbiology and Biotechnology
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• 제27권7호
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• pp.1233-1241
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• 2017

The ginsenoside Rh2 has strong anti-cancer, anti-inflammatory, and anti-diabetic effects. However, the application of ginsenoside Rh2 is restricted because of the small amounts found in Korean white and red ginsengs. To enhance the production of ginsenoside Rh2-MIX (comprising 20(S)-Rh2, 20(R)-Rh2, Rk2, and Rh3 as a 10-g unit) with high specificity, yield, and purity, a new combination of enzymatic conversion using the commercial enzyme Viscozyme L followed by acid treatment was developed. Viscozyme L treatment at pH 5.0 and $50^{\circ}C$ was used initially to transform the major ginsenosides Rb1, Rb2, Rc, and Rd into ginsenoside F2, followed by acid-heat treatment using citric acid 2% (w/v) at pH 2.0 and $121^{\circ}C$ for 15 min. Scale-up production in a 10-L jar fermenter, using 60 g of the protopanaxadiol-type ginsenoside mixture from ginseng roots, produced 24 g of ginsenoside Rh2-MIX. Using 2 g of Rh2-MIX, 131 mg of 20(S)-Rh2, 58 mg of 20(R)-Rh2, 47 mg of Rk2, and 26 mg of Rh3 were obtained at over 98% chromatographic purity. Then, the anti-cancer effect of the four purified ginsenosides was investigated on B16F10, MDA-MB-231, and HuH-7 cell lines. As a result, these four rare ginsenosides markedly inhibited the growth of the cancer cell lines. These results suggested that rare ginsenoside Rh2-MIX could be exploited to prepare an anti-cancer supplement in the functional food and pharmaceutical industries.