• Title/Summary/Keyword: BIOENGINEERING AND TECHNOLOGY

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Construction of an Efficient Mutant Strain of Trichosporonoides oedocephalis with HOG1 Gene Deletion for Production of Erythritol

  • Li, Liangzhi;Yang, Tianyi;Guo, Weiqiang;Ju, Xin;Hu, Cuiying;Tang, Bingyu;Fu, Jiaolong;Gu, Jingsheng;Zhang, Haiyang
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.700-709
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    • 2016
  • The mitogen-activated protein kinase HOG1 (high-osmolarity glycerol response pathway) plays a crucial role in the response of yeast to hyperosmotic shock. Trichosporonoides oedocephalis produces large amounts of polyols (e.g., erythritol and glycerol) in a culture medium. However, the effects of HOG1 gene knockout and environmental stress on the production of these polyols have not yet been studied. In this study, a To-HOG1 null mutation was constructed in T. oedocephalis using the loxP-Kan-loxP/Cre system as replacement of the targeted genes, and the resultant mutants showed much smaller colonies than the wild-type controls. Interestingly, compared with the wild-type strains, the results of shake-flask culture showed that To-HOG1 null mutation increased erythritol production by 1.44-fold while decreasing glycerol production by 71.23%. In addition, this study investigated the effects of citric acid stress on the T. oedocephalis HOG1 null mutants and the wild-type strain. When the supplementation of citric acid in the fermentation medium was controlled at 0.3% (w/v), the concentration of erythritol produced from the wild-type and To-HOG1 knockout mutant strains improved by 18.21% and 21.65%, respectively.

Laccase Production Using Pleurotus ostreatus 1804 Immobilized on PUF Cubes in Batch and Packed Bed Reactors: Influence of Culture Conditions

  • Prasad K. Krishna;Mohan S. Venkata;Bhaskar Y. Vijaya;Ramanaiah S. V.;Babu V. Lalit;Pati B. R.;Sarma P. N.
    • Journal of Microbiology
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    • v.43 no.3
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    • pp.301-307
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    • 2005
  • The feasibility of laccase production by immobilization of Pleurotus ostreatus 1804 on polyurethane foam (PUF) cubes with respect to media composition was studied in both batch and reactor systems. Enhanced laccase yield was evidenced due to immobilization. A relatively high maximum laccase activity of 312.6 U was observed with immobilized mycelia in shake flasks compared to the maximum laccase activity of free mycelia (272.2 U). It is evident from this study that the culture conditions studied, i.e. biomass level, pH, substrate concentration, yeast extract concentration, $Cu^{2+}$ concentration, and alcohol nature, showed significant influence on the laccase yield. Gel electrophoretic analysis showed the molecular weight of the laccase produced by immobilized P. ostreatus to be 66 kDa. The laccase yield was significantly higher and more rapid in the packed bed reactor than in the shake flask experiments. A maximum laccase yield of 392.9 U was observed within 144 h of the fermentation period with complete glucose depletion.

Effect of Light with Different Wavelengths on Nostoc flagelliforme Cells in Liquid Culture

  • Dai, Yu-Jie;Li, Jing;Wei, Shu-Mei;Chen, Nan;Xiao, Yu-Peng;Tan, Zhi-Lei;Jia, Shi-Ru;Yuan, Nan-Nan;Tan, Ning;Song, Yi-Jie
    • Journal of Microbiology and Biotechnology
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    • v.23 no.4
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    • pp.534-538
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    • 2013
  • The effects of lights with different wavelengths on the growth and the yield of extracellular polysaccharides of Nostoc flagelliforme cells were investigated in a liquid cultivation. N. flagelliforme cells were cultured for 16 days in 500 ml conical flasks containing BG11 culture medium under $27{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ of light intensity and $25^{\circ}C$ on a rotary shaker (140 rpm). The chlorophyll a, phycocyanin, allophycocyanin, and phycoerythrin contents in N. flagelliforme cells under the lights of different wavelengths were also measured. It was found that the cell biomass and the yield of polysaccharide changed with different wavelengths of light. The biomass and the yield of extracellular polysaccharides under the red or violet light were higher than those under other light colors. Chlorophyll a, phycocyanin, and allophycocyanin are the main pigments in N. flagelliforme cells. The results showed that N. flagelliforme, like other cyanobacteria, has the ability of adjusting the contents and relative ratio of its pigments with the light quality. As a conclusion, N. flagelliforme cells favor red and violet lights and perform the complementary chromatic adaptation ability to acclimate to the changes of the light quality in the environment.

Advances in the Early Detection of Lung Cancer using Analysis of Volatile Organic Compounds: From Imaging to Sensors

  • Li, Wang;Liu, Hong-Ying;Jia, Zi-Ru;Qiao, Pan-Pan;Pi, Xi-Tian;Chen, Jun;Deng, Lin-Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.11
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    • pp.4377-4384
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    • 2014
  • According to the World Health Organization (WHO), 1.37 million people died of lung cancer all around the world in 2008, occupying the first place in all cancer-related deaths. However, this number might be decreased if patients were detected earlier and treated appropriately. Unfortunately, traditional imaging techniques are not sufficiently satisfactory for early detection of lung cancer because of limitations. As one alternative, breath volatile organic compounds (VOCs) may reflect the biochemical status of the body and provide clues to some diseases including lung cancer at early stage. Early detection of lung cancer based on breath analysis is becoming more and more valued because it is non-invasive, sensitive, inexpensive and simple. In this review article, we analyze the limitations of traditional imaging techniques in the early detection of lung cancer, illustrate possible mechanisms of the production of VOCs in cancerous cells, present evidence that supports the detection of such disease using breath analysis, and summarize the advances in the study of E-noses based on gas sensitive sensors. In conclusion, the analysis of breath VOCs is a better choice for the early detection of lung cancer compared to imaging techniques. We recommend a more comprehensive technique that integrates the analysis of VOCs and non-VOCs in breath. In addition, VOCs in urine may also be a trend in research on the early detection of lung cancer.

Study of Macrophage Activation and Structural Characteristics of Purified Polysaccharide from the Fruiting Body of Cordyceps militaris

  • Lee, Jong-Seok;Kwon, Jeong-Seok;Won, Dong-Pil;Lee, Jung-Hyun;Lee, Keun-Eok;Lee, Shin-Young;Hong, Eock-Kee
    • Journal of Microbiology and Biotechnology
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    • v.20 no.7
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    • pp.1053-1060
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    • 2010
  • Cordyceps militaris, an entomopathogenic fungus belonging to the class Ascomycetes, has been reported to have beneficial biological activities such as hypoglycemic, anti-inflammatory, antitumor, antimetastatic, hypolipidemic, immunomodulatory, and antioxidant effects. In this study, the crude water-soluble polysaccharide CMP, which was obtained from the fruiting body of C. militaris by hot water extraction and ethanol precipitation, was fractionated by DEAE-cellulose and Sepharose CL-6B column chromatographies. This process resulted in three polysaccharide fractions, termed CMP Fr I, CMP Fr II, and CMP Fr III. Of these fractions, CMP Fr II, with an average molecular mass of 127 kDa, was able to upregulate effectively the phenotypic functions of macrophages such as NO production and cytokine expression. The chemical property of the stimulatory polysaccharide, CMP Fr II, was determined based on its monosaccharide composition, which consisted of glucose (56.4%), galactose (26.4%), and mannose (17.2%). Its structural characteristics were investigated by a combination of chemical and instrumental analyses, including methylation, reductive cleavage, acetylation, Fourier transform infrared spectroscopy (FTIR), and gas chromatography-mass spectrometry (GCMS). Results indicated that CMP Fr II consisted of the (1${\rightarrow}$4) or (1${\rightarrow}$2) linked glucopyranosyl or galactopyranosyl residue with a (1${\rightarrow}$2) or (1${\rightarrow}$6) linked mannopyranosyl, glucopyranosyl, or galactopyranosyl residue as a side chain. The configuration of the ${\beta}$-linkage and random coil conformation of CMP Fr II were confirmed using a Fungi-Fluor kit and Congo red reagent, respectively.

Xylitol Production by Kluyveromyces marxianus 36907-FMEL1 at High Temperature was Considerably Increased through the Optimization of Agitation Conditions (교반속도 최적화를 통한 Kluyveromyces marxianus 36907-FMEL1로 부터 고온에서의 자일리톨 생산성 증진)

  • Kim, Jin-Seong;Park, Jae-Bum;Jang, Seung-Won;Kwon, Deok-Ho;Hong, Eock Kee;Shin, Won Cheol;Ha, Suk-Jin
    • Microbiology and Biotechnology Letters
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    • v.45 no.1
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    • pp.57-62
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    • 2017
  • Recently, we isolated the mutant Kluyveromyces marxianus 36907-FMEL1, which demonstrated improved xylose reductase activity as compared to the parental strain, K. marxianus ATCC 36907. Effects of agitation conditions on xylitol production were verified using a bioreactor system. Under an agitation speed of 400 rpm, K. marxianus 36907-FMEL1 exhibited the highest xylitol yield (0.57 g/g) and productivity ($0.64g{\cdot}l^{-1}{\cdot}h^{-1}$) at $30^{\circ}C$. When the fermentation temperature was increased to $40^{\circ}C$, interestingly, xylitol yield and productivity were found to be increased to 21% (0.64 g/g) and 58% ($0.90g{\cdot}l^{-1}{\cdot}h^{-1}$), respectively, under the optimized agitation conditions.

Production of Human Interferon β by Recombinant E. coli Using the Codon Optimized Gene (코돈 최적화된 유전자를 이용한 재조합 대장균으로부터 인간 인터페론 베타 발현)

  • Kim, Jong-Seok;Jang, Seung-Won;Park, Jae-Bum;Kwon, Deok-Ho;Chang, Young-Jun;Jung, Hyung-Moo;Han, Sang-In;Hong, Eock-Kee;Ha, Suk-Jin
    • KSBB Journal
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    • v.32 no.1
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    • pp.16-21
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    • 2017
  • The multiple sclerosis caused by multiple inflammatory disease or immune system disorder, is usually treated by interferon ${\beta}$ through adjusting the abnormal immune reactions. For high production of human interferon ${\beta}$ using recombinant E. coli, codon optimized and wild type genes were synthesized. When pET-15b or pET-21a vector was used as an expression vector with each gene, there was no target protein expression. When pQE30 vector was used as an expression vector, human interferon ${\beta}$ was expressed by recombinant E. coli XL1-blue and E. coli JM109. Using the codon optimized gene, the expression of human interferon ${\beta}$ was slightly increased as compared to that from wild type gene. However, most of expressed human interferon ${\beta}$ was insoluble form.

The Application of Thermotolerant Yeast Kluyveromyces marxianus as a Potential Industrial Workhorse for Biofuel Production

  • Park, Jae-Bum;Kim, Jin-Seong;Jang, Seung-Won;Hong, Eunsoo;Ha, Suk-Jin
    • KSBB Journal
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    • v.30 no.3
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    • pp.125-131
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    • 2015
  • Kluyveromyces marxianus is a well-known thermotolerant yeast. Although Saccharomyces cerevisiae is the most commonly used yeast species for ethanol production, the thermotolerant K. marxianus is more suitable for simultaneous saccharification and fermentation (SSF) processes. This is because enzymatic saccharification usually requires a higher temperature than that needed for the optimum growth of S. cerevisiae. In this study, we compared the fermentation patterns of S. cerevisiae and K. marxianus under various temperatures of fermentation. The results show that at a fermentation temperature of $45^{\circ}C$, K. marxianus exhibited more than two fold higher growth rate and ethanol production rate in comparison to S. cerevisiae. For SSF using starch or corn stover as the sole carbon source by K. marxianus, the high temperature ($45^{\circ}C$) fermentations showed higher enzymatic activities and ethanol production compared to SSF at $30^{\circ}C$. These results demonstrate the potential of the thermotolerant yeast K. marxianus for SSF in the industrial production of biofuels.