• KSBB Journal
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• 제22권1호
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• pp.37-42
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• 2007

본 연구에서는 배양시간에 따른 Thraustochytrium aureum ATCC 34304 균체 내부에 저장하는 지질의 지방산조성 및 균체 막에 축척하는 지질 내의 지방산조성의 변화 과정을 정량적으로 규명하였다. 건조균체량은 배양 5일째에 3.94 g/L를 나타내었으며, 균체당의 지질 함량은 배양 3일째에 최대값 25.0%를 나타내었다. 균체 내의 triacylglyceride (TAG)량은 배양 3일까지는 증가하였으나 그 이후에는 거의 일정한 값을 유지하였다. 반면 세포막을 구성하는 phospholipid (PL)의 함량은 3일까지는 감소하나 그 이후에는 일정한 값을 나타내었다. TAG를 구성하고 있는 다중불포화지방산의 함량은 배양 초기 60.3%이였으나 배양 5일 후에는 45.3%까지 감소하였으며, DHA는 42.1%에서 33.9% 까지 감소하였다. 포화지방산의 함량은 배양 초기에는 24.9%이었으나 배양 5일 후에는 27.8%까지 증가하였다. PL 내의 불포화지방산의 함량은 배양시간에 따라 48.0%에서 17.5%까지 크게 감소하는 경향을 나타내었으나, 포화지방산의 함량은 오차범위 내에서의 감소 경향을 나타내었다. 본 연구 결과에 의하면 배양시간이 경과하여 배양액내의 영양 환경이 열악하게 되는 경우 균주는 포화지방산보다 불포화지방산을 에너지원으로 우선 사용한다고 해석할 수 있다.

• 한국식품과학회지
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• 제41권6호
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• pp.694-699
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• 2009

프로폴리스 에탄올 추출물의 항돌연변이 및 항균효과를 시험하여 국내산과 미국산 프로폴리스의 생리활성을 평가하였다. 항돌연변이 활성은 Ames test로, 항균활성은 여드름 형성에 관여하는 미생물인 P. acnes, S. Epidermidis, S. aureus, P. aeruginosa에 대한 생육 억제효과를 paper disk 법과 agar dilution 법으로 평가하였다. S. Typhimurium TA98 균주에 있어서 $1-200\;{\mu}g$/plate 농도로 70% 에탄올 추출물을 첨가했을 때 Trp-p-1에 의해 유도된 돌연변이에 대해 국내산 0.9-91.3%, 미국산 0.5-104.2%, 2-AA에 의해 유도된 돌연변이에 대해서는 국내산 37.6-94.4%, 미국산 21.0-97.7%의 돌연변이 억제효과를 용량 의존적으로 각각 나타내었다. 항균활성을 paper disk 법으로 평가했을 때 P. acnes, S. Epidermidis, S. aureus, P. aeruginosa에 대해 70% 에탄올 추출물의 $5,000\;{\mu}g$/paper disk 농도에서의 생육 저지환의 크기는 국내산이 각각 5.7, 5.9, 6.8, 5.2 mm, 미국산이 각각 5.7, 6.2, 6.7, 5.9 mm이었다. Agar dilution 법으로 평가한 MIC는 P. acnes, P. aeruginosa S. Epidermidis, S. aureus, P. aeruginosa에 대해 국내산, 미국산 모두 5,000, 1,500, 1,500, $1,500\;{\mu}g/mL$로 나타났다.

• KSBB Journal
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• 제24권6호
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• pp.495-507
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• 2009

21세기에 들어서면서 해양에 대한 인식은 해양선진국들을 중심으로 하루가 다르게 변모하고 있으며, 석유 및 육상자원의 고갈의 가속화로 인하여 해양의 중요성은 점차 커져 가고 있다. 지속가능한 발전을 위해서는 해조류, 해수 등과 같은 해양자원으로부터 산업용 화학품, 신소재, 연료, 희귀금속 등의 생산을 가능하게 하는 해양화학생물산업의 개발이 절실히 필요해지고 있다. 본 글에서는 해양화학생물산업분야 중 세계적으로 각광 받고 있으며 우리나라에서도 발전 가능성이 큰 분야인 해조다당류산업, 해양신소재산업, 해양바이오연료산업, 해양제염산업, 해양심층수산업 등 다섯분야를 선정하고 이들 분야의 현황과 전망을 소개하였다. 각 분야별 소개는 산업발전 역사, 기술개발 수준, 산업화 정도를 포함하고 있으며, 국내외 산업경향과 앞으로의 발전 가능성을 조사하였다. 분석결과, 해양화학생물산업기술의 발전을 위해서는 해양생물공정 개발에 대한 적극적인 연구개발 투자와 해양생물과학자들과 생물화학공학자들간의 협력 연구가 앞으로 더 많이 필요하기는 하지만 해양화학생물산업의 미래전망은 매우 밝은 것으로 나타났다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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• pp.482-493
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• 2005

Biomass is originally photosynthesized from inorgainic compounds such as $CO_2$, minerals, water and solar energy. Recent studies have shown that anaerobic bacteria have the ability to convert recalcitrant biomass such as cellullosic or chitinoic materials to useful compounds. The biomass containing agricultural waste, unutilized wood and other garbage is expected to utilize as feed, food and fuel by microbial degradation and other metabolic functions. In this study we isolated several anaerobic, cellulolytic and chitinolytic bacteria from rumen fluid, compost and soil to study their related enzymes and genes. The anaerobic and cellulolytic bacteria, Clostridium thermocellum, Clostridium stercorarium, and Clostridium josui, were isolated from compost and the chitinolytic Clostridium paraputrificum from beach soil and Ruminococcus albus was isolated from cow rumen. After isolation, novel cellulase and xylanase genes from these anaerobes were cloned and expressed in Escherichia coli. The properties of the cloned enzymes showed that some of them were the components of the enzyme (cellulase) complex, i.e., cellulosome, which is known to form complexes by binding cohesin domains on the cellulase integrating protein (Cip: or core protein) and dockerin domains on the enzymes. Several dockerin and cohesin polypeptides were independently produced by E. coli and their binding properties were specified with BIAcore by measuring surface plasmon resonance. Three pairs of cohesin-dockerin with differing binding specificities were selected. Two of their genes encoding their respective cohesin polypeptides were combined to one gene and expressed in E. coli as a chimeric core protein, on which two dockerin-dehydrogenase chimeras, the dockerin-formaldehyde dehydrogenase and the dockerin-NADH dehydrogenase are planning to bind for catalyzing $CO_2$ reduction to formic acid by feeding NADH. This reaction may represent a novel strategy for the reduction of the green house gases. Enzymes from the anaerobes were also expressed in tobacco and rice plants. The activity of a xylanase from C. stercorarium was detected in leaves, stems, and rice grain under the control of CaMV35S promoter. The digestibility of transgenic rice leaves in goat rumen was slightly accelerated. C. paraputrificum was found to solubilize shrimp shells and chitin to generate hydrogen gas. Hydrogen productivity (1.7 mol $H_2/mol$ glucos) of the organism was improved up to 1.8 times by additional expression of the own hydrogenase gene in C. paraputrficum using a modified vector of Clostridiu, perfringens. The hydrygen producing microflora from soil, garbage and dried pelletted garbage, known as refuse derived fuel(RDF), were also found to be effective in converting biomass waste to hydrogen gas.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권1호
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• pp.52-59
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• 2005

The study was targeted to saccharify foodwastes with the cellulolytic and amylolytic enzymes obtained from culture supernatant of Trichoderma harzianum FJ1 and analyze the kinetics of the saccharification in order to enlarge the utilization in industrial application. T. harzianum FJ1 highly produced various cellulolytic (filter paperase 0.9, carboxymethyl cellulase 22.0, ${\beta}$-glucosidase 1.2, Avicelase 0.4, xylanase 30.8, as U/mL-supernatant) and amylolytic (${alpha}$-amylase 5.6, ${\beta}$-amylase 3.1, glucoamylase 2.6, as U/mL-supernatant) enzymes. The $23{\sim}98\;g/L$ of reducing sugars were obtained under various experimental conditions by changing FPase to between $0.2{\sim}0.6\;U/mL$ and foodwastes between $5{\sim}20\%$ (w/v), with fixed conditions at $50^{\circ}C$, pH 5.0, and 100 rpm for 24 h. As the enzymatic hydrolysis of foodwastes were performed in a heterogeneous solid-liquid reaction system, it was significantly influenced by enzyme and substrate concentrations used, where the pH and temperature were fixed at their experimental optima of 5.0 and $50^{\circ}C$, respectively. An empirical model was employed to simplify the kinetics of the saccharification reaction. The reducing sugars concentration (X, g/L) in the saccharification reaction was expressed by a power curve ($X=K{\cdot}t^n$) for the reaction time (t), where the coefficient, K and n. were related to functions of the enzymes concentrations (E) and foodwastes concentrations (S), as follow: $K=10.894{\cdot}Ln(E{\cdot}S^2)-56.768,\;n=0.0608{\cdot}(E/S)^{-0.2130}$. The kinetic developed to analyze the effective saccharification of foodwastes composed of complex organic compounds could adequately explain the cases under various saccharification conditions. The kinetics results would be available for reducing sugars production processes, with the reducing sugars obtained at a lower cost can be used as carbon and energy sources in various fermentation industries.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권1호
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• pp.67-72
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• 2005

To achieve demineralization of crab shell waste by chemical and biological treatments, lactic acid and lactic acid bacterium were applied. In 5.0 and $10\%$ lactic acid, pH rapidly decreased from 6.8 to 4.2 and from 4.5 to 2.4 at day 3, respectively, and thereafter the pH remained at an almost constant level. In a $10\%$ lactic acid bacterium inoculum, pH lowered to 4.6 at day 5. Relative residual ash content rapidly decreased to 49.1 and $16.4\%$ in 5 and $10\%$ lactic acid treatments, respectively, for the initial 12 h. In 2.5, 5 and $10\%$ lactic acid bacterium inoculums, relative residual ash content rapidly decreased to 55.2, 40.9 and $44.7\%$, respectively, on the first day. Residual dry masses were 76.4, 67.8 and $46.6\%$ in 2.5, 5 and $10\%$ lactic acid treatments, respectively, for the initial 12 h. After a one-time exchange of the lactic acid solution, in the $5.0\%$ lactic acid treatment, residual dry mass rapidly decreased from 66.0 to $41.4\%$. In 2.5, 5 and $10\%$ lactic acid bacterium inoculums, residual dry masses decreased to 67.6, 57.4 and $59.6\%$ respectively, on the first day. Protein contents after demineralization ranged from $51.3{\sim}54.7\%$ in the chemical treatments and decreased to $32.3\%$ in the lactic acid fermentation process. A negative relationship was shown between pH and demineralization rate in lactic acid and lactic acid bacterium treatments. These results suggest that lactic acid fermentation can be an alternative for demineralization of crab shells, even though the rate and efficiency of the demineralization is lower than the chemical treatment.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권2호
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• pp.127-133
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• 2006

Shewanella oneidensis MR-1 has the ability to inhale certain metals and chemical compounds and exhale these materials in an altered state; as a result, this microorganism has been widely applied in bioremediation protocols. However, the relevant characteristics of cell growth and biosynthesis of PuFAs have yet to be thoroughly investigated. Therefore, in this study, we have attempted to characterize the growth and fatty acid profiles of S. oneidensis MR-1 under a variety of temperature conditions. The fastest growth of S. oneidensis MR-1 was observed at $30^{\circ}C$, with a specific growth rate and doubling time of $0.6885h^{-1}\;and\;1.007 h$. The maximum cell mass of this microorganism was elicited at a temperature of $4^{\circ}C$. The eicosapentaenoic acid (EPA) synthesis of S. oneidensis MR-1 was evaluated under these different culture temperatures. S. oneidensis MR-1 was found not to synthesize EPA at temperatures in excess of $30^{\circ}C$, but was shown to synthesize EPA at temperatures below $30^{\circ}C$. The EPA content was found to increase with decreases in temperature. We then evaluated the EPA biosynthetic pathway, using a phylogenetic tree predicted on 16s rRNA sequences, and the homology of ORFs between S. oneidensis MR-1 and Shewanella putrefaciens SCRC-2738, which is known to harbor a polyketide synthase (PKS)-like module. The phylogenetic tree revealed that MR-1 was very closely related to both Moritella sp., which is known to synthesize DHA via a PKS-like pathway, and S. putrefaciens, which has been reported to synthesize EPA via an identical pathway. The homology between the PKS-like module of S. putrefaciens SCRC-2738 and the entire genome of S. oneidensis MR-1 was also analyzed, in order to mine the genes associated with the PKS-like pathway in S. oneidensis MR-1. A putative PKS-like module for EPA biosynthesis was verified by this analysis, and was also corroborated by the experimental finding that S. oneidensis MR-1 was able to synthesize EPA without the expression of $dihomo-{\gamma}-linoleic$ acid (DGLA) and arachidonic acid (AA) formed during EPA synthesis via the FAS pathway.

• Asian-Australasian Journal of Animal Sciences
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• 제26권4호
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• pp.537-544
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• 2013

This study examined whether pre-treating palm kernel expeller (PKE) with exogenous enzyme would degrade its fiber content; thus improving its metabolizable energy (ME), growth performance, villus height and digesta viscosity in broiler chickens fed diets containing PKE. Our results showed that enzyme treatment decreased (p<0.05) hemicellulose and cellulose contents of PKE by 26.26 and 32.62%, respectively; and improved true ME (TME) and its nitrogen corrected value ($TME_n$) by 38% and 33%, respectively, compared to the raw sample. Average daily gain (ADG), feed intake and feed conversion ratio (FCR) of chickens fed on different dietary treatments in the grower period were not significantly different. Although there was no difference in feed intake (p>0.05) among treatment groups in the finisher period, ADG of chickens in the control (PKE-free diet) was higher (p<0.05) than in all treatment groups fed either 20 or 30% PKE, irrespective of with or without enzyme treatment. However, ADG of birds fed with 20% PKE was higher than those fed with 30% PKE. The FCR of chickens in the control was the lowest (2.20) but not significantly different from those fed 20% PKE diets while birds in the 30% PKE diets recorded higher (p>0.05) FCR. The intestinal villus height and crypt depth (duodenum, jejunum and ileum) were not different (p>0.05) among treatments except for duodenal crypt depth. The villus height and crypt depth of birds in enzyme treated PKE diets were higher (p<0.05) than those in the raw PKE groups. Viscosity of the intestinal digesta was not different (p>0.05) among treatments. Results of this study suggest that exogenous enzyme is effective in hydrolyzing the fiber (hemicellulose and cellulose) component and improved the ME values of PKE, however, the above positive effects were not reflected in the growth performance in broiler chickens fed the enzyme treated PKE compared to those received raw PKE. The results suggest that PKE can be included up to 5% in the grower diet and 20% in the finisher diet without any significant negative effect on FCR in broiler chickens.

• 한국식품영양과학회지
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• 제44권12호
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• pp.1905-1911
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• 2015

천연 미백소재 개발과 관련하여 많은 연구들이 멜라닌 합성저해 및 활성 메커니즘을 규명하는 데 초점이 맞춰졌으며, 이러한 이유로 tyrosinase 저해제 개발이 다양하게 이루어져 왔다. 본 연구는 배 유래의 polyphenol oxidase를 이용하여 천연에 존재하는 단순 폴리페놀인 pyrogallol의 산화 축합반응을 유도하여 purpurogallin을 효율적으로 생합성하였으며, 본 화합물에 대해서 미백 활성을 평가하였다. 먼저 MTT assay를 통해 세포독성이 없는 농도구간을 설정하였으며, purpurogallin은 $25{\mu}M$ 농도의 melanoma 세포 내에서 tyrosinase 활성을 20% 이상 저해하는 것을 확인하였다. 또한 $25{\mu}M$의 시험 농도에서 purpurogallin은 약 20% 이상의 melanin 생합성 저해 활성을 나타내었다. 미백 관련 전사인자인 MITF, TRP-1, TRP-2, tyrosinase의 단백질 발현을 측정한 결과, 본 화합물은 B16F10 melanoma 세포에서 tyrosinase, TRP-1과 TRP-2의 단백질 생합성을 두 추출물 모두 억제하는 것을 확인하였다. Tyrosinase, TRP-1과 TRP-2의 발현을 조절하는 전사인자로는 MITF가 관여하는 것으로 알려져 있으며, 실제로 MITF는 melanin 생성과 관련된 여러 유전자의 발현을 조절하는 데 중요한 작용을 하고 있다. 따라서 purpurogallin은 melanin 생성과 관련된 중요한 세 가지 단백질의 생합성을 전사단계에서 조절 전사인자인 MITF의 단백질 발현을 억제하는 효과가 있음을 확인하였다. 이상의 결과로부터 멜라닌 생합성에 있어서 상위 신호단계에 있는 전사인자 MITF의 활성을 억제함으로써 하위 신호전달 과정을 억제하는 것임을 시사하며, 향후 추가적인 검증작업을 통해 화장품 소재화가 가능할 것으로 판단된다.

• 생명과학회지
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• 제27권5호
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• pp.567-574
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• 2017

본 연구는 막걸리로부터 ${\gamma}$-amino butyric acid (GABA) 생성 유산균을 분리 및 동정하고 최적 GABA 생산조건을 확립하는데 그 목적이 있다. 막걸리로부터 64균주의 유산균은 MRS 배지에서 성장된 집락의 색과 모양의 특성에 따라 분리하였다. 분리균주의 GABA 생산은 1% MSG가 첨가된 MRS 액체배지에서 배양하여 TLC와 HPLC 방법에 의해 평가되었다. B-134 균주는 GABA생성을 위한 우수균주로 선발하였다. 16S rRNA 유전자 및 glutamate decarboxylase B (gadB) 유전자의 염기서열분석을 통하여, B-134 균주는 Lactobacillus plantarum subsp. plantarum B-134 균주로 명명하였다. GABA 생성을 위한 온도, pH, NaCl 및 MSG 농도를 달리하여 최적배양조건을 조사하였다. 그 결과 B-134 균주의 최적배양 조건은 온도 $37^{\circ}C$, pH 5.7, NaCl 농도 0% (w/v), 그리고 MSG 농도 3% (w/v)로 결정되었으며 본 조건에서 48시간 배양시 25 mM의 GABA를 생산하였다. 이러한 결과로부터 B-134 균주는 GABA함유 건강기능식품개발을 위한 유용한 균주로 판단된다.