• Journal of Microbiology and Biotechnology
• /
• 제16권6호
• /
• pp.911-920
• /
• 2006

[ $CD8^+$ ] T Iymphocytes with the cytotoxic activity and capability to release various cytokines are the major players in immune responses against viral infection and cancer. To identify the proteins specific to resting or activated human CD8$^+$ T cells, human CD8$^+$ T cells were activated with anti-CD3+anti-CD28 mAb in the presence of IL-2. The solubilized proteins from resting and activated human CD8$^+$ T cells were separated by high-resolution two-dimensional polyacrylamide gel electrophoresis, and their proteomes were analyzed. Proteomic analysis of resting and activated T cells resulted in identification of 35 proteins with the altered expression. Mass spectrometry coupled with Profound and SWISS-PROT database analysis revealed that these identified proteins are to be functionally associated with cell proliferation, metabolic pathways, antigen presentation, and intracellular signal transduction pathways. We also identified six unknown proteins predicted from genomic DNA sequences specific to resting or activated CD8$^+$ T cells. Protein network studies and functional characterization of these novel proteins may provide new insight into the signaling transduction pathway of CD8$^+$ T cell activation.

• Journal of Microbiology and Biotechnology
• /
• 제25권6호
• /
• pp.837-844
• /
• 2015

Carob waste is a useful raw material for the second-generation ethanol because 50% of its dry weight is sucrose, glucose, and fructose. To optimize the process, we have studied the influence of the initial concentration of sugars on the fermentation performance of Saccharomyces cerevisiae. With initial sugar concentrations (S₀ ) of 20 g/l, the yeasts were derepressed and the ethanol produced during the exponential phase was consumed in a diauxic phase. The rate of ethanol consumption decreased with increasing S₀ and disappeared at 250 g/l when the Crabtree effect was complete and almost all the sugar consumed was transformed into ethanol with a yield factor of 0.42 g/g. Sucrose hydrolysis was delayed at high S₀ because of glucose repression of invertase synthesis, which was triggered at concentrations above 40 g/l. At S₀ higher than 250 g/l, even when glucose had been exhausted, sucrose was hydrolyzed very slowly, probably due to an inhibition at this low water activity. Although with lower metabolic rates and longer times of fermentation, 250 g/l is considered the optimal initial concentration because it avoids the diauxic consumption of ethanol and maintains enough invertase activity to consume all the sucrose, and also avoids the inhibitions due to lower water activities at higher S₀ .

• Journal of Microbiology and Biotechnology
• /
• 제27권8호
• /
• pp.1379-1385
• /
• 2017

The content of taxol in the bark of yews is very low, and this is not affordable from the environmental point of view. Thus, it is a necessity to look for alternative sources of taxol production to solve its supply. Currently, a large portion of the taxol in the market comes from chemical semi-synthesis, but the semi-synthetic precursors such as baccatin III and 10-deacetyl-baccatin III are extracted from needles and twigs of yew trees. Taxol-producing fungi as a renewable resource is a very promising way to increase the scale of taxol production. Our group has obtained a taxol-producing endophytic fungus, Aspergillus niger subsp. taxi HD86-9, to examine if A. niger can produce the taxanes. Six compounds from the fermentation broth of strain HD86-9 were isolated and identified by $^1H$ NMR, $^{13}C$ NMR, and ESI-MS. The results showed that the six compounds included four taxane diterpenoids (taxol, cephalomannine, baccatin III, and 10-deacetyl-baccatin III) and two non-taxane compounds (${\beta}-sitosterol$ and flavonoid isovitexin). The study verified that the taxanes can be produced by the A. niger, which is very important to taxol production via chemical semi-synthesis. Additionally, the finding is potentially very significant to solve the taxol semi-synthetic precursors extracted from needles and twigs of yew trees, and the precursor production can be easily increased through the culture condition optimization, genetic breeding, and metabolic engineering of the A. niger.

• 한국환경성돌연변이발암원학회지
• /
• 제8권1호
• /
• pp.13-21
• /
• 1988

Mutagenic actions of aflatoxin B$_1$ (AFB$_1$) in the presence of various concentrations of L-ascorbIc acid (AA) in Salmonella typhimurium strains TA 100 and TA98 were studied. Spontaneous revertants per plate of the tester strains TA100 and TA98 were 121-125 and 25-30 with or without S9 mix, respectively. The negative controls used in the study did not show any mutagenesis in the tester strains. AFB$_1$ revealed strong mutagenicity at the dose levels of 0.05, 0.1 and 0.25 ${\mu}$g/plate with metabolic activation system in both strains. However, it showed a toxic effect when the levels were more than 0.5 ${\mu}$g/plate. When lower concentrations of AA (5-20 ${\mu}$g/plate) were added to AFB$_1$ in the Ames assay system with S9 mix the mutagenic action of AFB$_1$ decreased in both strains. About 70-90% of mutagenicity of AFB$_1$ disappeared in strain TA100 when 20${\mu}$g of AA was added to 0.05 ${\mu}$g of AFB$_1$. The inhibitory effect was greatly increased by the addition of higher concentrations of AA to AFB$_1$ in TA100 strain. The mutagenicity of AFB$_1$ was completely inhibited when 100 ${\mu}$g and 500 ${\mu}$g of AA were added to 0.05 ${\mu}$g and 0.1 ${\mu}$g of AFB$_1$, respectively, However, this protective effect of AA on AFB$_1$ mediated mutagenesis was less effective in TA98 strain than that in TA100.

• BMB Reports
• /
• 제48권9호
• /
• pp.501-506
• /
• 2015

Based on the potential beneficial effects of growth hormone releasing peptide (GHRP)-6 on muscle functions, a newly synthesized GHRP-6-biotin conjugate was tested on cultured myoblast cells. Increased expression of myogenic marker proteins was observed in GHRP-6-biotin conjugate-treated cells. Additionally, increased expression levels of insulin-like growth factor-1 and collagen type I were observed. Furthermore, GHRP-6-biotin conjugate-treated cells showed increased metabolic activity, as indicated by increased concentrations of energy metabolites, such as ATP and lactate, and increased enzymatic activity of lactate dehydrogenase and creatine kinase. Finally, binding protein analysis suggested few candidate proteins, including desmin, actin, and zinc finger protein 691 as potential targets for GHRP6-biotin conjugate action. These results suggest that the newly synthesized GHRP-6-biotin conjugate has myogenic stimulating activity through, at least in part, by stimulating collagen type I synthesis and several key proteins. Practical applications of the GHRP-6-biotin conjugate could include improving muscle condition. [BMB Reports 2015; 48(9): 501-506]

• BMB Reports
• /
• 제47권1호
• /
• pp.27-32
• /
• 2014

Plant abiotic stress tolerance has been modulated by engineering the trehalose synthesis pathway. However, many stress-tolerant plants that have been genetically engineered for the trehalose synthesis pathway also show abnormal development. The metabolic intermediate trehalose 6-phosphate has the potential to cause aberrations in growth. To avoid growth inhibition by trehalose 6-phosphate, we used a gene that encodes a bifunctional in-frame fusion (BvMTSH) of maltooligosyltrehalose synthase (BvMTS) and maltooligosyltrehalose trehalohydrolase (BvMTH) from the nonpathogenic bacterium Brevibacterium helvolum. BvMTS converts maltooligosaccharides into maltooligosyltrehalose and BvMTH releases trehalose. Transgenic rice plants that over-express BvMTSH under the control of the constitutive rice cytochrome c promoter (101MTSH) or the ABA-inducible Ai promoter (105MTSH) show enhanced drought tolerance without growth inhibition. Moreover, 101MTSH and 105MTSH showed an ABA-hyposensitive phenotype in the roots. Our results suggest that over-expression of BvMTSH enhances drought-stress tolerance without any abnormal growth and showes ABA hyposensitive phenotype in the roots.

• Molecules and Cells
• /
• 제25권2호
• /
• pp.231-241
• /
• 2008

Indole glucosinolates (IG) play important roles in plant defense, plant-insect interactions, and stress responses in plants. In an attempt to metabolically engineer the IG pathway flux in Chinese cabbage, three important Arabidopsis cDNAs, CYP79B2, CYP79B3, and CYP83B1, were introduced into Chinese cabbage by Agrobacterium-mediated transformation. Overexpression of CYP79B3 or CYP83B1 did not affect IG accumulation levels, and overexpression of CYP79B2 or CYP79B3 prevented the transformed callus from being regenerated, displaying the phenotype of indole-3-acetic acid (IAA) overproduction. However, when CYP83B1 was overexpressed together with CYP79B2 and/or CYP79B3, the transformed calli were regenerated into whole plants that accumulated higher levels of glucobrassicin, 4-hydroxy glucobrassicin, and 4-methoxy glucobrassicin than wild-type controls. This result suggests that the flux in Chinese cabbage is predominantly channeled into IAA biosynthesis so that coordinate expression of the two consecutive enzymes is needed to divert the flux into IG biosynthesis. With regard to IG accumulation, overexpression of all three cDNAs was no better than overexpression of the two cDNAs. The content of neoglucobrassicin remained unchanged in all transgenic plants. Although glucobrassicin was most directly affected by overexpression of the transgenes, elevated levels of the parent IG, glucobrassicin, were not always accompanied by increases in 4-hydroxy and 4-methoxy glucobrassicin. However, one transgenic line producing about 8-fold increased glucobrassicin also accumulated at least 2.5 fold more 4-hydroxy and 4-methoxy glucobrassicin. This implies that a large glucobrassicin pool exceeding some threshold level drives the flux into the side chain modification pathway. Aliphatic glucosinolate content was not affected in any of the transgenic plants.

• 대한핵의학회지
• /
• 제38권2호
• /
• pp.161-170
• /
• 2004

분자영상은 살아있는 개체의 몸 속에서 일어나는 생물학적 반응이나 특정한 표적분자를 비관혈적이며 반복적으로 영상화하는 기술이다. 이를 위해서는 두 가지 기본 요소가 요구되는 바 하나는 관심 생물현상에 의해 농도나 분광특성이 변하는 분자영상용 추적자이며 다른 하나는 이런 추적자를 모니터링하는 장비이다. 분자 핵의학 영상기술은 이제 신경과학분야에서도 활발히 적용되고 있으며 신경관련 기초연구나 뇌질환 관련 신약개발에 이미 중요한 역할을 하고 있다. 최근에는 살아있는 개체에서 약제 투여가 뇌에 미치는 약물학적, 생리적 영향을 조사하는 데에도 이용되고 있다. 다가오는 미래에는 각종 뇌질환에서 특이적 표적을 공략하는 새로운 분자치료가 개발되어 뇌질환 치료에 혁명적인 변화를 가져올 것으로 예상되고 있다. 그 예로, 파킨슨씨 병과 같은 퇴행성 신경질환에 줄기세포를 이용한 자가수선, 신경보호, 약물분비 치료, 성장인자와의 병행치료 등이 개발되고, 유전자 치료도 이용될 것으로 보인다. 신경 분자 핵의학 영상은 이와 같은 새로운 뇌질환 치료기술의 개발에 있어서 뇌 안에서 일어나는 분자수준의 변화를 실시간으로 모니터링함으로써 관련연구에 크게 기여할 것으로 기대된다.

• Advances in Energy Research
• /
• 제2권1호
• /
• pp.1-9
• /
• 2014

The green microalgae Chlamydomonas reinhardtti is well-known specie in the terms of $H_2$ production by photo fermentation and has been studying for a long time. Although the $H_2$ production yield is promising; there are some bottlenecks to enhance the yield and efficiency to focus on a well-designed, sustainable production and also scaling up for further studies. D1 protein of photosystem II (PSII) plays an important role in photosystem damage repair and related to $H_2$ production. Because Chlamydomonas is the model algae and the genetic basis is well-studied; metabolic engineering tools are intended to use for enhanced production. Mutations are focused on D1 protein which aims long-lasting hydrogen production by blocking the PSII repair system thus $O_2$ sensitive hydrogenases catalysis hydrogen production for a longer period of time under anaerobic and sulfur deprived conditions. Chlamydomonas CC124 as control strain and D1 mutant strains(D240, D239-40 and D240-41)are cultured photomixotrophically at $80{\mu}mol\;photons\;m^{-2}s^{-1}$, by two sides. Cells are grown in TAP medium as aerobic stage for culture growth; in logarithmic phase cells are transferred from aerobic to an anaerobic and sulfur deprived TAP- S medium and 12 mg/L initial chlorophyll content for $H_2$ production which is monitored by the water columns and later detected by Gas Chromatography. Total produced hydrogen was $82{\pm}10$, $180{\pm}20$, $196{\pm}20$, $290{\pm}30mL$ for CC124, D240, D239-40, D240-41, respectively. $H_2$ production rates for mutant strains was $1.3{\pm}0.5mL/L.h$ meanwhile CC124 showed 2-3 fold lower rate as $0.57{\pm}0.2mL/L.h$. Hydrogen production period was $5{\pm}2days$ for CC124 and mutants showed a longer production time for $9{\pm}2days$. It is seen from the results that $H_2$ productions for mutant strains have a significant effect in terms of productivity, yield and production time.

• Journal of Plant Biotechnology
• /
• 제36권3호
• /
• pp.207-213
• /
• 2009

Starch serves not only as an energy source for plants, animals, and humans but also as an environmentally friendly alternative for fossil fuels. Progress in understanding of starch biosynthesis, and the isolation of many genes involved in this process have enabled the genetic modification of crops in a rational manner to produce novel starches with improved functionality. Starch is composed of two glucose polymers, amylose and amylopectin. The amylose and amylopectin ratio in starch affects its physical and physicochemical properties. Alteration in starch structure can be achieved by modifying genes encoding the enzymes responsible for starch biosynthesis and starch hydrolysis. Here, we describe recent findings concerning the starch modification in sweetpotato. Sweetpotato [Ipomoea batatas (L.) Lam] ranks seventh in annual production among food crops in the world as an important starch source. To develop transgenic sweetpotato plants with modifying starch composition, we constructed transformation vectors overexpressing granule bound starch synthase I and inhibiting amylopectin synthesis genes such as starch branching enzyme and isoamylase under the control of 35S promoter, respectively. Transformation of sweetpotato (cv. Yulmi) is in progress.