• Journal of Microbiology and Biotechnology
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• 제25권7호
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• pp.1056-1069
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• 2015

Osmotic pressure is a critical factor for erythritol production with osmophilic yeast. Protein expression patterns of an erythritol-producing yeast, Yarrowia lipolytica, were analyzed to identify differentially-expressed proteins in response to osmotic pressure. In order to analyze intracellular protein levels quantitatively, two-dimensional gel electrophoresis was performed to separate and visualize the differential expression of the intracellular proteins extracted from Y. lipolytica cultured under low (3.17 osmol/kg) and high (4.21 osmol/kg) osmotic pressures. Proteomic analyses allowed identification of 54 differentially-expressed proteins among the proteins distributed in the range of pI 3-10 and 14.4-97.4 kDa molecular mass between the osmotic stress conditions. Remarkably, the main proteins were involved in the pathway of energy, metabolism, cell rescue, and stress response. The expression of such enzymes related to protein and nucleotide biosynthesis was inhibited drastically, reflecting the growth arrest of Y. lipolytica under hyperosmotic stress. The improvement of erythritol production under high osmotic stress was due to the significant induction of a range of crucial enzymes related to polyols biosynthesis, such as transketolase and triosephosphate isomerase, and the osmotic stress responsive proteins like pyridoxine-4-dehydrogenase and the AKRs family. The polyols biosynthesis was really related to an osmotic response and a protection mechanism against hyperosmotic stress in Y. lipolytica. Additionally, the high osmotic stress could also induce other cell stress responses as with heat shock and oxidation stress responses, and these responsive proteins, such as the HSPs family, catalase T, and superoxide dismutase, also had drastically increased expression levels under hyperosmotic pressure.

• Journal of Microbiology and Biotechnology
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• 제24권4호
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• pp.497-506
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• 2014

In this study, crude glycerol was used as a carbon source in the cultivation of wild yeasts, aiming at the production of microbial lipids and citric acid. Forty yeasts of different sources were tested concerning their growth in crude and commercial glycerol. Four yeasts (Lindnera saturnus UFLA CES-Y677, Yarrowia lipolytica UFLA CM-Y9.4, Rhodotorula glutinis NCYC 2439, and Cryptococcus curvatus NCYC 476) were then selected owing to their ability to grow in pure ($OD_{600}$ 2.133, 1.633, 2.055, and 2.049, respectively) and crude ($OD_{600}$ 2.354, 1.753, 2.316, and 2.281, respectively) glycerol (10%, 20%, and 30%). Y. lipolytica UFLA CM-Y9.4 was selected for its ability to maintain cell viability in concentrations of 30% of crude glycerol, and high glycerol intake (18.907 g/l). This yeast was submitted to lipid production in 30 g/l of crude glycerol, and therefore obtained 63.4% of microbial lipids. In the fatty acid profile, there was a predominance of stearic (C18:0) and palmitic (C16:0) acids in the concentrations of 87.64% and 74.67%, respectively. We also performed optimization of the parameters for the production of citric acid, which yielded a production of 0.19 g/l of citric acid in optimum conditions (38.4 g/l of crude glycerol, agitation of 184 rpm, and temperature of $30^{\circ}C$). Yarrowia lipolytica UFLA CM-Y9.4 presented good lipid production when in the concentration of 30 g/l of glycerol. These data may be used for production in large quantities for the application of industrial biodiesel.

• 한국균학회지
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• 제48권2호
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• pp.161-167
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• 2020

국내에 자생하는 지렁이로(붉은줄지렁이)의 장에서 야생효모를 분리, 동정하고자 하였다. 지렁이는 고양시 난지 물 재생 센터에서 분양 받았으며, 지렁이 5마리에서 19개의 야생효모를 분리하였다. 효모의 D1/D2 영역으로 동정하여 국내에 보고되지 않은 효모 3균주를 선별하였다. 국내 미기록종 효모인 Yarrowia deformans YP242 (=KACC48778), Sporidiobolus pararoseus YP66(=KCTC27963), Naganishia liquefaciens YI9 (=KACC48948) 균주는 위상차현미경, API 20C AUX kit를 통해 균학적 특성을 조사하였다. YP242, YI9, YP66 균주의 세포 모양은 타원형이며, 집락은 볼록하고, 부드러운 재질을 가지는 것으로 관찰되었다. 탄소원 활성 측정 결과, YP242는 glycerol, L-arabinose, N-acetyl-D-glucosamine을, YP66는 N-acetyl-D-glucosamine을, liquefaciens YI9는 2-keto-D-gluconate을 탄소원으로 사용할 수 있음을 확인하였다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.144-144
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• 2005

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권5호
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• pp.320-326
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• 2000

Promoters of the genes G3P, ICL1, POT1, POX1, POX2 and POX5 of the yeast Y. lipolytica were studied in respect to their regulations and activities during growth on different carbon sources. The aim of this study was to select suitable promoters for high expression of heterologous genes in this yeast. For this purpose the promoters were fused with the reporter gene lacZ of E. coli and integrated as single copies into the genome of Y. lipolytica strain PO1d. The measurement of expressed activities of ${\beta}$-galactosidase revealed that pICL1, pPOX2 and pPOT1 are the strongest regulable promoters available for Y. lipolytica, at present. pPOX2 and pPOT1 were highly induced during growth on oleic acid and were completely repressed by glucose and glycerol. pICL1 was strongly inducible by ethanol besides alkanes and fatty acids, however, not completely repressible by glucose or glycerol. Ricinoleic acid methyl ester appeared as a very strong inducer for pPOT1 and pPOX2, in spite of that it inhibited growth of Y. lipolytica transformants.

• 한국미생물·생명공학회지
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• 제40권2호
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• pp.98-103
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• 2012

경기도 일대에서 수집한 메주 시료에서 지질분해 활성을 나타내는 균주 Y124를 분리하여 동정한 결과 Yarrowia lipolytica와 100% 상동성을 보였다. 분리 균주가 생산하는 lipase의 조효소에 대한 일반적인 특성을 조사한 결과, 탄소원으로 olive oil을 단독으로 사용한 YPO 배지에서 8시간 배양하였을 때 lipase 활성이 가장 높게 나타났다. YPD 배지에서는 lipase 활성이 거의 없었으며, olive oil과 glucose를 모두 포함하는 YPDO 배지에서는 lipase 활성이 YPO 배지 보다 낮았다. 그리고 olive oil 농도에 따른 lipase 활성을 측정한 결과, olive oil 무첨가보다 0.7% 첨가하여 8시간 배양했을 때 lipase 활성이134 U/mL으로 가장 높게 나타나 lipase의 생산이 olive oil의 첨가에 의해 유도되는 것으로 생각된다. 생육온도에 따른 lipase 활성 측정한 결과, $30^{\circ}C$에 배양하였을 때 배양 8시간에 가장 높은 활성이 나타났고, $25^{\circ}C$와 $37^{\circ}C$에 배양하였을 때는 배양 12시간에 활성이 가장 높게 나타났으며, Y124균주의 lipase 활성 최적 온도는 $30^{\circ}C$로 나타났다. 그리고 lipase의 기질 친화도를 확인한 결과 Y124균주가 생산하는 lipase의 경우 p-nitrophenyl octanoate ($C_8$)에서 가장 높은 활성이 나타났다.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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• pp.1563-1570
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• 2010

To construct a highly sensitive detection system for endocrine disruptors (EDs), we have compared the activity of promoters with the n-alkane-inducible cytochrome P450 gene (ALK1), isocitrate lyase gene (ICL1), ribosomal protein S7 gene (RPS7), and the translation elongation factor-1${\alpha}$ gene (TEF1) for the heterologous gene in Yarrowia lipolytica. The promoters were introduced into the upstream of the lacZ or hERa reporter genes, respectively, and the activity was evaluated by ${\beta}$-galactosidase assay for lacZ and Western blot analysis for hER${\alpha}$. The expression analysis revealed that the ALK1 and ICL1 promoters were induced by n-decane and by EtOH, respectively. The constitutive promoter of RPS7 and TEF1 showed mostly a high level of expression in the presence of glucose and glycerol, respectively. In particular, the TEF1 promoter showed the highest ${\beta}$-galactosidase activity and a significant signal by Western blotting with the anti-estrogen receptor, compared with the other promoters. Moreover, the detection system was constructed with promoters linked to the upstream of the expression vector for the hER${\alpha}$ gene transformed into the Y. lipolytica with a chromosome-integrated lacZ reporter gene under the control of estrogen response elements (EREs). It was indicated that a combination of pTEF1p-hER${\alpha}$ and CXAU1-2XERE was the most effective system for the $E_2$-dependent induction of the ${\beta}$-galactosidase activity. This system showed the highest ${\beta}$-galactosidase activity at $10^{-6}\;M\;E_2$, and the activity could be detected at even the concentration of $10^{-10}\;M\;E_2$. As a result, we have constructed a strongly sensitive detection system with Y. lipolitica to evaluate recognized/suspected ED chemicals, such as natural/synthetic hormones, pesticides, and commercial chemicals. The results demonstrate the utility, sensitivity, and reproducibility of the system for identifying and characterizing environmental estrogens.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2005년도 International Meeting of the Microbiological Society of Korea
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• pp.203.4-203
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• 2005

• Journal of Microbiology and Biotechnology
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• 제29권1호
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• pp.141-150
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• 2019

The spatial landmark protein Bud8 plays a crucial role in bipolar budding in the budding yeast Saccharomyces cerevisiae. The unconventional yeast Yarrowia lipolytica can also bud in a bipolar pattern, but is evolutionarily distant from S. cerevisiae. It encodes the protein YALI0F12738p, which shares the highest amino acid sequence homology with S. cerevisiae Bud8, sharing a conserved transmembrane domain at the C-terminus. Therefore, we named it YlBud8. Deletion of YlBud8 in Y. lipolytica causes cellular separation defects, resulting in budded cells remaining linked with one another as cell chains or multiple buds from a single cell, which suggests that YlBud8 may play an important role in cell separation, which is distinct from the function of Bud8 in S. cerevisiae. We also show that the YlBud8-GFP fusion protein is located at the cell membrane and enriched in the bud cortex, which would be consistent with a role in the regulation of cell separation. The coiled-coil domain at the N-terminus of YlBud8 is important to the correct localization and function of YlBud8, as truncated proteins that do not contain the coiled-coil domain cannot rescue the defects observed in $Ylbud8{\Delta}$. This finding suggests that a new signaling pathway controlled by YlBud8 via regulation of cell separation may exist in Y. lipolytica.

• Journal of Microbiology and Biotechnology
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• 제8권6호
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• pp.719-722
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• 1998

The addition of decane to biotransfonnation media containing Yarrowia lipolytica led to the accumulation of intennediate L-phenylacetaldehyde and L-phenethyl acetate during bioconversion of L-phenylalanine, whilst none of these products were obtained in conventional aqueous fennentations. The results obtained support an earlier hypothesis (Spinnler et al. 1996. Proc. Natl. A cad. Sci. USA 93: 3373-3376) that organic solvents, acting as "thermodynamic traps" for hydrophobic intermediates, can substantially alter metabolic fluxes.