• 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.

• 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.

• 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.

• 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.

• Applied Biological Chemistry
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• v.38 no.1
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• pp.26-30
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• 1995

To produce ethanol from Jerusalem artichoke powder efficiently, Kluyveromyces marxianus FO43 cells were encapsulated in 2% sodium alginate and were cultured in batch reactor to investigate the fermentation properties. Batch culture of immobilized cells left for 4 days in 15% Jerusalem artichoke medium showed ethanol concentration of 3.38%(w/v) and ethanol yield to theoretical value of 54.20%, lower than 3.76%(w/v) and 71.13% for the culture of free cells. Addition of cellulase to $15{\sim}20%$ Jerusalem artichoke media increased the production of ethanol, owing to remarkable reduction in consistency of the suspension. So it was possible to achieve an ethanol concentration of 5.57%(w/v) arid an ethanol yield to theoretical value of 68.86% in even 20% Jerusalem artichoke medium by cultivation of immobilized cells for 4 days. The alginate beads showed constant ethanol productivity after recycling 11 times (22 days) in repeated batch fermentation.

• Microbiology and Biotechnology Letters
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• v.25 no.4
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• pp.386-390
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• 1997

The characteristics of alcohol dehydrogenase (ADH, EC 1.1.1.1, alcohol:NAD oxidoreductase) of thermotolerant alcohol-producing yeasts, Saccharomyces cerevisiae RA-74-2 and Kluyveromyces marxianus RA-912, were compared with that of mesophilic S. cerevisiae D, an industrial strain. Under anaerobic culture condition, both S. cerevisiae RA-74-2 and D had similar level of ADH activity at 30$\circ$C, and the activity of S. cerevisiae RA-74-2 at 37$\circ$C was the same level at 30$\circ$C. However, the level of ADH activity of S. cerevisiae D at 37$\circ$C decreased about 70% of that at 30$\circ$C. The level of enzyme activity of K. marxianus RA-912, which showed lower alcohol productivity than S. cerevisiae RA-74-2 and D, was about 43% of those strains at 30$\circ$C, and decreased somewhat at 37$\circ$C. The results showed a good correlation between the alcohol productivities and the level of ADH activities of these strains grown at 30$\circ$C and 37$\circ$C. And the higher heat stability of ADH of S. cerevisiae RA-74-2 than that of S. cerevisiae D seemed to reflect the ability of high temperature fermentation. Despite of its fermentation ability even at 45$\circ$C, however, the ADH of K. marxianus RA-912 showed lower heat stability than that of S. cerevisiae D. Both S. cerevisiae RA-74-2 and D showed similar patterns of two bands of ADH isozyme, and the low band of S. cerevisiae RA-74-2 moved slightly faster than that of S. cerevisiae D. The staining intensity of the bands of S. cerevisiae D at 37$\circ$C was weaker than those at 30$\circ$C. However, S. cerevisiae RA-74-2 showed no differences in total intensity of the bands of 30$\circ$C and 37$\circ$C. As the patterns of cellular proteins and ADH isozyme of K. marxianus RA-912 were different from S. cerevisiae RA-74-2 and D, K. marxianus might have its own characteristic ADH system.

• Microbiology and Biotechnology Letters
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• v.19 no.3
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• pp.265-271
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• 1991

In order to produce alcohol for the alternative energy from dried powder of Jerusalem artichoke was investigated with Kluyveromyces marxianus UCD(FST)55-82, which was reported to assimilate inulin. The optimal condition for the production of ethanol by K. marxianus was elucidated to be incubation temperature of $30^{\circ}C$, initial pH 5.44, agitation of 100 rpm, 1,000 ml of medium in a 2.5l-vessel, anaerobic state, and inoculation of 2.5%(v/v). Addition of antifoam A concentrate(si1icon polymer) of 0.01% and urea of 0.1% increased the concentration of ethanol effectively. The optimized condition showed ethanol concentration of 6.8%(v/v) in Jerusalem artichoke liquid medium, production yield of 91.91% and productivity of 2.71 g/l/hr during the first day and 0.71g ethanol/l/hr during four days of incubation.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1529-1533
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• 2010

The inulinase gene (INU1) from Kluyveromyces marxianus NCYC2887 was overexpressed by using the GAL10 promotor in a ${\Delta}ga180$ strain of Saccharomyces cerevisiae. The inulinase gene lacking the original signal sequence was fused in-frame to a mating factor ${\alpha}$ signal sequence for secretory expression. Use of the ${\Delta}ga180$ strain allowed for the galactose-free induction of inulinase expression using a glucose-only medium. Shake-flask cultivation in YPD medium produced 34.6 U/ml of the recombinant inulinase, which was approximately 13-fold higher than that produced by K. marxianus NCYC2887. It was found that the use of the ${\Delta}ga180$ strain improved the expression of inulinase in the recombinant S. cerevisiae in both aerobic and anaerobic conditions by about 2.9- and 1.7-fold, respectively. A 5-l fed-batch fermentation using YPD medium was performed under aerobic condition with glucose feeding, which resulted in the inulinase production of 31.7 U/ml at the $OD_{600}$ of 67. Ethanol fermentation of dried powder of Jerusalem artichoke, an inulin-rich biomass, was also performed using the recombinant S. cerevisiae expressing INU1 and K. marxianus NCYC2887. Fermentation in a 5-l scale fermentor was carried out at an aeration rate of 0.2 vvm, an agitation rate of 300 rpm, and with the pH controlled at 5.0. The temperature was maintained at $30^{\circ}C$ and $37^{\circ}C$, respectively, for the recombinant S. cerevisiae and K. marxianus. The maximum productivities of ethanol were 59.0 and 53.5 g/l, respectively.

• Journal of Microbiology and Biotechnology
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• v.26 no.7
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• pp.1259-1266
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• 2016

Bioethanol was produced from Kappaphycus alvarezii seaweed biomass using separate hydrolysis and fermentation (SHF). Pretreatment was evaluated for 60 min at 121℃ using 12% (w/v) biomass slurry with 364 mM H₂SO₄. Enzymatic saccharification was then carried out at 45℃ for 48 h using Celluclast 1.5 L. Ethanol fermentation with 12% (w/v) K. alvarezii hydrolyzate was performed using the yeasts Saccharomyces cerevisiae KCTC1126, Kluyveromyces marxianus KCTC7150, and Candida lusitaniae ATCC42720 with or without prior adaptation to high concentrations of galactose. When non-adapted S. cerevisiae, K. marxianus, and C. lusitaniae were used, 11.5 g/l, 6.7 g/l, and 6.0 g/l of ethanol were produced, respectively. When adapted S. cerevisiae, K. marxianus, and C. lusitaniae were used, 15.8 g/l, 11.6 g/l, and 13.4 g/l of ethanol were obtained, respectively. The highest ethanol concentration was 15.8 g/l, with Y_EtOH = 0.43 and Y_T% = 84.3%, which was obtained using adapted S. cerevisiae.

• 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.