• Title/Summary/Keyword: Kluyveromyces marxianus

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Overexpression of Mutant Galactose Permease (ScGal2_N376F) Effective for Utilization of Glucose/Xylose or Glucose/Galactose Mixture by Engineered Kluyveromyces marxianus

  • Kwon, Deok-Ho;Kim, Saet-Byeol;Park, Jae-Bum;Ha, Suk-Jin
    • Journal of Microbiology and Biotechnology
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    • v.30 no.12
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    • pp.1944-1949
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    • 2020
  • Mutant sugar transporter ScGAL2-N376F was overexpressed in Kluyveromyces marxianus for efficient utilization of xylose, which is one of the main components of cellulosic biomass. K. marxianus ScGal2_N376F, the ScGAL2-N376F-overexpressing strain, exhibited 47.04 g/l of xylose consumption and 26.55 g/l of xylitol production, as compared to the parental strain (24.68 g/l and 7.03 g/l, respectively) when xylose was used as the sole carbon source. When a mixture of glucose and xylose was used as the carbon source, xylose consumption and xylitol production rates were improved by 195% and 360%, respectively, by K. marxianus ScGal2_N376F. Moreover, the glucose consumption rate was improved by 27% as compared to that in the parental strain. Overexpression of both wild-type ScGAL2 and mutant ScGAL2-N376F showed 48% and 52% enhanced sugar consumption and ethanol production rates, respectively, when a mixture of glucose and galactose was used as the carbon source, which is the main component of marine biomass. As shown in this study, ScGAL2-N376F overexpression can be applied for the efficient production of biofuels or biochemicals from cellulosic or marine biomass.

Increased Production of Exoinulinase in Saccharomyces cerevisiae by Expressing the Kluyveromyces marxianus INU1 Gene Under the Control of the INU1 Promoter

  • KIM HEE EUN;QIN RUI;CHAE KEON SANG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.447-450
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    • 2005
  • Aspergillus niger inuE gene and Kluyveromyces marxianus INUI gene coding for exoinulinase were expressed in Saccharomyces cerevisiae under the control of K. marxianus INUI promoter. Recombinant S. cerevisiae expressing K. marxianus exoinulinase produced maximum 85 U/ml into culture medium, which was 9- to 14-fold higher than the activity produced by any other strain reported so far. In addition, K. marxianus INUI promoter produced 20- fold higher activity than S. cerevisiae glyceraldehydes phosphate dehydrogenase (GPD) promoter in S. cerevisiae.

Quality characteristics of whey Makgeolli by Kluyveromyces marxianus (Kluyveromyces marxianus에 의한 유청막걸리 품질특성)

  • Kim, Su-Hwan;Huh, Chang-Ki;Kim, So-Mang;Cho, In-Kyung;Kim, Yong-Doo
    • Food Science and Preservation
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    • v.22 no.4
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    • pp.528-534
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    • 2015
  • This study was performed to analyze the processing and quality characteristics of whey Makgeolli prepared with different types of yeast and ratio of the whey content. Lactose content of yeast culture medium containing S. cerevisiae. was 1.36% whereas lactose content of yeast culture containing K. marxianus KCCM 12015 was very little. Yeast culture of both K. marxianus KCCM 35455 and K. marxianus KCCM 50700 did not produce lactose. Until the 10th day, ethanol production ability of S. cerevisiae, K. marxianus KCCM 12015, K. marxianus KCCM 35455, and K. marxianus KCCM were 0.31%, 2.51%, 2.53%, and 2.59%, respectively. Total acids content increased rapidly with the increase in the addition of whey content in the initial 2 days and then decreased during 4~10 days of fermentation. In the aspect of pH, the pH was rapidly decreased in the initial 2 days and then increased until 10th day of fermentation with the increase in whey content. Ethanol content of whey Makgeollis at 10th day of fermentation was the highest in yeast culture containing K. marxianus. From the sensory evaluation, the flavor score of whey Makgeollisin was higher than that of control. The color and taste scores were increased as the increase in the addition of whey. The comprehensive preference indicated that Makgeolli prepared with 100% whey was the best among other samples.

Immobilization of Kluyveromyces marxianus FO43 for Ethanol Production (Kluyveromyces marxianus FO43의 Algiante 고정화와 에탄올 발효특성)

  • Lee, Hee-Suk;Shin, Ji-Hyun;Choi, Eon-Ho
    • Applied Biological Chemistry
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    • v.38 no.1
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    • pp.20-25
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    • 1995
  • This experiment was attempted to improve ethanol productivity by immobilization of Kluyveromyces marxianus FO43 using Jerusalem artichoke powder. Sucrose medium was used to determine optimum conditions for cell immobilization. The optimum conditions were alginate concentration of 2%, bead size of 2 mm, a particle input ratio of 30 : 100, cultivation period of 24 hours, and substrate concentration of 10%(w/v). The immobilized cells produced the high concentrations of ethanol at pH $4.5{\sim}6.5$ and $30{\sim}45^{\circ}C$, broader ranges of pH and temperatures than those of free cells. Under optimum conditions the immobilized cells showed ethanol concentration of 46.4 g/L and productivity of 1.93 g/L.h. The microphotograph using a two phase contrast microscope showed that immobilized cells cultivated under the optimum conditions were densely populated toward the surface area of beads.

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Characteristics of Functional Fermented Milk by Mixed Starters of Lactobacillus bulgaricus and Kluyveromyces marxianus (Lactobacillus bulgaricus와 Kluyveromyces marxianus의 혼합 스타터를 이용한 기능성 발효유의 특성)

  • Yoon Won-Ho;Nam Bo-Ra;Kim Jin-Man;Kim Chang-Han
    • Food Science of Animal Resources
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    • v.26 no.2
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    • pp.252-256
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    • 2006
  • This study was carried out to investigate characteristics of acid and alcohol fermented milk by mixed starters made by Lactobacillus bulgaricus (KCTC 3635) and Kluyveromyces marxianus (KCTC 17212) for 36 hours when the curds were formed. Final pH and titratable acidity were about 4.5 and 0.68%, respectively. The viable cell counts of lactic acid bacteria and yeast for alcohol fermented milk were increased to $3.2{\times}10^9 CFU/mL$ and $5.3{\times}10^9 CFU/mL$, respectively. The ethanol contents increased to 0.35% during fermentation. Antitumor activities of the fermented milk against tumor cell lines, such as HEp-2, HEC-1B, SW-156 and SK-MES-1 showed to 86.6, 70.3, 60.4 and 57.14%, respectively.

Sulfuric Acid Hydrolysis and Detoxification of Red Alga Pterocladiella capillacea for Bioethanol Fermentation with Thermotolerant Yeast Kluyveromyces marxianus

  • Wu, Chien-Hui;Chien, Wei-Chen;Chou, Han-Kai;Yang, Jungwoo;Lin, Hong-Ting Victor
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1245-1253
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    • 2014
  • One-step sulfuric acid saccharification of the red alga Pterocladiella capillacea was optimized, and various detoxification methods (neutralization, overliming, and electrodialysis) of the acid hydrolysate were evaluated for fermentation with the thermotolerant yeast Kluyveromyces marxianus. A proximate composition analysis indicated that P. capillacea was rich in carbohydrates. A significant galactose recovery of $81.1{\pm}5%$ was also achieved under the conditions of a 12% (w/v) biomass load, 5% (v/v) sulfuric acid, $121^{\circ}C$, and hydrolysis for 30 min. Among the various detoxification methods, electrodialysis was identified as the most suitable for fermentable sugar recovery and organic acid removal (100% reduction of formic and levulinic acids), even though it failed to reduce the amount of the inhibitor 5-HMF. As a result, K. marxianus fermentation with the electrodialyzed acid hydrolysate of P. capillacea resulted in the best ethanol levels and fermentation efficiency.

Impact of sodium or potassium concentration in glucose aquoes solution to fermentation by Kluyveromyces marxianus (배양액내 나트륨과 칼륨의 농도가 고온 발효 균주 Kluyveromyces marxianus의 발효에 미치는 영향)

  • Song, Woo-Yong;Shin, Soo-Jeong
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.47 no.3
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    • pp.11-17
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    • 2015
  • In acid hydrolysis process of biomass saccharification. neutralization of acid hydrolyzate is essential step, which resulted in dissolved cations in glucose solution. Impact of cations to Kluyveromyces marxianus in glucose solution was investigated focused on ethanol fermentation. Either potassium or sodium cations decreased the ethanol fermentation and glucose to ethanol conversion. Glucose consumption by K. marxianus was delayed by increasing potassium cation concentration as completely consumed within 12 h in potassium cation 0.46 mol and 0.92 mol but within 24 h in potassium cation 1.38 mol. Also, ethanol fermentation process was slowed down with increasing concentration of the potassium sulfate. Fermentation of glucose solution to ethanol was more inhibited by sodium cation than potassium cation in glucose solution. Glucose was completely consumed within 24 h in sodium cation 0.95 mol. but at 1.90 mol or 2.84 mol in sodium cation could not finish the fermentation within 48 hour. Ethanol concentration was 22.26 g/L at low sodium cation in glucose solution with complete fermentation within 24 h. With increasing sodium cation in glucose solution, final ethanol concentration was reached at 14.10 g/L (sodium cation con) and 0.21 g/L (sodium cation con), which meant delaying of fermentation by sodium cations.

Effect of Ethanol and/or Organic Acids on the Growth of Lactobacillus plantarum, Leuconostoc mesenteroides, Kluyveromyces marxianus Identified from Mul-kimchi (물김치 서식 미생물, Lactobacillus plantarum, Leuconostoc mesenteroides, Kluyveromyces marxianus 생육에 Ethanol과 유기산이 미치는 영향)

  • 한영숙
    • Journal of the East Asian Society of Dietary Life
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    • v.13 no.5
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    • pp.425-432
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    • 2003
  • Effects of ethanol and/or some organic acid on the growth of some microorganism grown in Kimchi, Lactobacillus plantarum, Leuconostoc mesenteroides. Kluyveromyces marxianus, were identified and the cell injury was observed by measuring optical density at 260nm. When 0.0∼5.0% ethanol was added in the growth medium, the cell growth was inhibited depending on the concentration. Organic acids involving acetic, adipic and citric acid inhibited the growth of L. plantarum and Leu. mesenteroides, but K. marxianus, the yeast, was not at 0.1% of organic acid. When 2.0% of ethanol and 0.1% of organic acid were used, adipic acid was more effective on the growth inhibition. This inhibition of microbial growth seemed to be caused by the leakage of internal contents from microbes which were observed by the optical density at 260nm in the buffer supernatant. 5.0% of ethanol accelerated the optical density increase at 260nm in L. plantarum, Leu. mesenteroides and K. marxianus, but 2.0% of ethanol did not. 0.1% organic acid increased the absorbance of the supernatant in lactic bacteria, but not in yeast, K. marxianus. The measurement of absorbance at 260nm revealed that the cell injury increased when 2.0% of ethanol and/or 0.1% of organic acid. especially adipic acid were added.

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Partial Purification and Characterization of Exoinulinase from Kluyveromyces marxianus YS-1 for Preparation of High-Fructose Syrup

  • Singh, Ram Sarup;Dhaliwal, Rajesh;Puri, Munish
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.733-738
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    • 2007
  • An extracellular exoinulinase($2,1-\beta-D$ fructan fructanohydrolase, EC 3.2.1.7), which catalyzes the hydrolysis of inulin into fructose and glucose, was purified 23.5-fold by ethanol precipitation, followed by Sephadex G-100 gel permeation from a cell-free extract of Kluyveromyces marxianus YS-1. The partially purified enzyme exhibited considerable activity between pH 5 to 6, with an optimum pH of 5.5, while it remained stable(100%) for 3 h at the optimum temperature of $50^{\circ}C$. $Mn^{2+}\;and\;Ca^{2+}$ produced a 2A-fold and 1.2-fold enhancement in enzyme activity, whereas $Hg^{2+}\;and\;Ag^{2+}$ completely inhibited the inulinase. A preparation of the partially purified enzyme effectively hydrolyzed inulin, sucrose, and raffinose, yet no activity was found with starch, lactose, and maltose. The enzyme preparation was then successfully used to hydrolyze pure inulin and raw inulin from Asparagus racemosus for the preparation of a high-fructose syrup. In a batch system, the exoinulinase hydrolyzed 84.8% of the pure inulin and 86.7% of the raw Asparagus racemosus inulin, where fructose represented 43.6mg/ml and 41.3mg/ml, respectively.

Purification and Characterization of Intracellular and Extracellular Inulase from Kluyveromyces marxianus (Kluyveromyces marxianus 가 생산하는 Intracellular 및 Extracellular Inulase 의 정제 및 특성비교)

  • Kim, Su-Il;Moon, Hang-Sik
    • Applied Biological Chemistry
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    • v.30 no.2
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    • pp.169-178
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    • 1987
  • The extracellular and intracellular inulases from Kluyveromyces marxianus were purified and characterized. The maximum production of both inulases was achieved at stationary phase in a pH-controlled medium at pH 5 with yeast nitrogen base as organic nitrogen source. Each enzyme was concentrated by tannic acid precipitation and separated into two fractions by DEAF-cellulose chromatography. Electrophoretic analysis showed that the four fractions had three glycoprotein bards each. Only main glycoprotein band, however, had both inulase and invertase activities. There were no significant differences between two enzymes in the optimum pH and temperature. But the intracellular inulases had higher heat stability and less affinity toward inulin than the extracellular enzymes do. All the purified enzymes were considered to be exo-inulases using hydrolyzate analysis with TLC.

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