• KSBB Journal
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• v.7 no.1
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• pp.1-7
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• 1992

Degradation of structural carbohydrates has been observed in samples of sweet sorghum inoculated with either Clostridium cellulolyticum or Bacteroides succinogenes. However, conditions under which these rellulolytic organisms can compete effectively with lactic acid bacteria have not yet been determined. Degradation of cellulose by B. succinogenes was found not to be inhibited by either glucose or succinate.

• Microbiology and Biotechnology Letters
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• v.19 no.4
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• pp.319-324
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• 1991

Three strains able to efficiently produce ethanol from cellulosic hydrolysates were isolated from soil samples by enrichment culture in liquid saccharified wheat bran medium. The profiles of physiological and biochemical properties of two yeasts KM-09 and KM-402 and a bacterium Hg-225 were almost identical from those of Candida sp. and Klebsiella sp., respectively. Strains KM-09 and HG-225 used xylose and cellobiose as fermentable sugars, and HG-225 had a wide range of sugar utilization for ethanol fermentation. The optimal pH and temperature for growth of KM-09, KM-402 and HG-225 were 5.8, 5.6 and 6.8 and 32t, $30^{\circ}C$~ and $38^{\circ}C$, respectively. During the ethanol fermentation in saccharified wheat bran by the isolated strains, optimal temperature for ethanol production was more or less higher than those for growth, and addition of 0.2% (w/v) $MgSO_4$, into the medium enhanced ethanol productivity. Of the three strains ethanol content of KM-09 was the highest with about 2.3% (v/v), and ethanol production rate of HG-225 was faster than the others and maximum productivity was after 4 days. KM-09 (1.42% v/v) and HG-225 (1.05%, vlv) produced ethanol from 4% (wIv) xylose but growth rate was slower than on glucose. Otherwise KM-402 showed the highest ethanol productivity on glucose, but no ethanol was detected on xylose and cellobiose.

• The Korean Journal of Mycology
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• v.24 no.3 s.78
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• pp.206-213
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• 1996

Talaromyces luteus 2004, a thermophilic mutant of T. luteus 6112 was obtained by mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine. T. luteus 2004 produced thermophilic carboxymethylcellulase (CMCase), and other polysaccharide enzymes: avicellase, xylanase, and ${\beta}-glucosidase$. Induction of CMCase production was shown at the highest level in 3% carboxymethylcellulose (CMC) minimal broth, indicating that CMC could work as an inducer. However, glucose and D-cellobiose showed catabolite repression for CMCase production which was under the control of CMC utilization. Optimal conditions for CMCase activity were at $70^{\circ}C$ and pH 4.0, suggesting that CMCase of T. luteus 2004 was a thermophilic enzyme.

• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.504-511
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• 2016

Lactulose, a synthetic disaccharide, has received increasing interest because of its role as a prebiotic that can increase the proliferation of Bifidobacterium and Lactobacillus spp. and enhance the absorption of calcium and magnesium. While the industrial production of lactulose is still mainly achieved by the chemical isomerization of lactose in alkaline media, this process has drawbacks including the need to remove catalysts and by-products, as well as high energy requirements. Recently, the use of cellobiose 2-epimerase (CE) has been considered an interesting alternative for industrial lactulose production. In this study, to develop a process for enzymatic lactulose production using CE, we screened improved mutant enzymes ($CS-H^RC^E$) from a library generated by an error-prone PCR technique. The thermostability of one mutant was enhanced, conferring stability up to $75^{\circ}C$, and its lactulose conversion yield was increased by 1.3-fold compared with that of wild-type CE. Using a recombinant Escherichia coli strain harboring a CS35 $H^RC^E$-expressing plasmid, we prepared cell beads immobilized on a Ca-alginate substrate and optimized their reaction conditions. In a batch reaction with 200 g/l lactose solution and the immobilized cell beads, lactose was converted into lactulose with a conversion yield of 43% in 2 h. In a repeated 38-plex batch reaction, the immobilized cell beads were relatively stable, and 80% of the original enzyme activity was retained after 4 cycles. In conclusion, we developed a reasonable method for lactulose production by immobilizing cells expressing thermostable CE. Further development is required to apply this approach at an industrial scale.

• Journal of Life Science
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• v.26 no.12
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• pp.1477-1486
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• 2016

Lactulose (4-O-${\beta}$-D-galactopyranosyl-D-fructose) is a non-digestible synthetic ketose disaccharide which can used in food and pharmaceutical fields due to its useful functions for encephalopathy, chronic constipation, hyperammonemia, etc. Therefore, the lactulose is regarded as one of the most important disaccharides and have been concentrated much interesting as an attractive functional material in the current industry. From this reason, the research related on the production of lactulose has been carried out various academic and industrial research groups. To produce lactulose, two main methods, chemical production and enzymatic production have been used. Commercially lactulose produced by alkaline isomerization of lactose as chemical production method but it has many disadvantages such as rapid lactulose degradation, purification, and waste management. From these reasons, lactulose produced by enzymatic method which solves these problems has been suggested as a proper method for lactulose production. Two different enzymatic methods have been reported as methods for lactulose production. Lactulose can be obtained through hydrolysis and transfer reaction catalyzed by a ${\beta}$-galactosidase which requires fructose as co-substrate and exhibits a low conversion. Alternatively, lactulose can be produced by direct isomerization of lactose to lactulose catalyzed by cellobiose 2-epimerase which requires lactose as a single substrate and achieves a high lactulose yield. This review summarizes the current state of lactulose production by chemical and biological methods.

• Polymer(Korea)
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• v.32 no.3
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• pp.230-238
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• 2008

The thermal and optical properties of cellobiose octa(cholestryloxycarbonyl)alkanoates CCCBn, $n=2{\sim}8$,10, the number of methylene units in the spacer) were investigated. All the samples formed monotropic cholesteric phases with left-handed helical structures. CCBn with n=2 or 10, in contrast with CCBn with $3{\leq}n{\leq}8$, did not display reflection colors over the full cholesteric range, suggesting that the helical twisting power of the cholesteryl group highly depends on the length of the spacer connecting the cholesteryl group to the cellobiose chain. The isotropic-cholestropic transition ($T_{ic}$) and glass transition temperatures decreased with increasing n and showed no odd-even effect. The transition entropy at $T_{ic}$ increased with increasing n from 2 up 6, but at n=7 it drops significantly and then increased again with increasing n from 8 to 10. The sharp change at n=7 may be attributed to a difference in arrangement of the side groups. The thermal stability and degree of order in the mesophase and the temperature dependence of the optical pitch observed for CCBn were significantly different from those reported for the cellulose tri(cholesteryloxycarbonyl)alkanoates and glucose penta(cholesteryloxycarbonyl)alkanoates. The results were discussed in terms of the differences in the degree of polymerization, the number of the mesogenic units per mole-glucose unit, and the conformation of the molecules.

• KSBB Journal
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• v.7 no.4
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• pp.265-270
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• 1992

Low cost fermentation substrates frequently contain a mixture of carbon sources including hexoses, pentoses and disaccharides. Fermentation of such mixtures requires an understanding of how each of these substrates is utilized. During batch culture of Pichia stipitis CBS 5776 on sugar mixtures, glucose causes catabolite repression of xylose and cellobiose utilization. Also, glucose causes a permanent repression of xylose utilization as evidenced by reduced growth rates during the xylose phase of glucose/xylose fermentation. The growth model for multiple substrates is developed based on a cyclic AMP mediated catabolite repression mechanism and this model adequately described the growth and ethanol production from sugar mixtures.

• Microbiology and Biotechnology Letters
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• v.24 no.4
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• pp.385-392
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• 1996

Bacillus stearotheymophilus, a potent xylanolytic bacterium isolated from soil, was tested for the strain's strategies of pentose utilization and the evidence of substrate preferences. The strain metabolized glucose, xylose, ribose, maltose, cellobiose, sucrose, arabinose and xylitol. The efficacy of the sugars as a carbon and energy source in this strain was of the order named above. The organism, however, could not grow on glycerol as a sole growth substrate. During cultivation on a mixture of glucose and xylose or arabinose, the major hydrolytic products of xylan, B. stearothermophilus displayed classical diauxic growth in which glucose was utilized during the first phase. On the other hand, the pentose utilization was prevented immediately upon addition of glucose. Cellobiose was preferred over xylose or arabinose. In contrast, maltose and pentose were co-utilized, and also no preference on between xylose and arabinose. Enzymatic studies indicated that B. stearothermophilus possessed constitutive hexokinase, a key enzyme of the glucose metabolic system. While, the production of $^{D}$-xylose isomerase, $^{D}$-xylulokinase and $^{D}$-arabinose isomerase essential for pentose phosphate pathway were induced by xylose, xylan, and xylitol but repressed by glucose. Taken together, the results suggested that the sequential utilization of B. stearothermophilus would be mediated by catabolite regulatory mechanisms such as catabolite inhibition or inducer exclusion.

• Microbiology and Biotechnology Letters
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• v.23 no.6
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• pp.652-658
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• 1995

We attempted simultaneous expression of genes coding for endoglucanase and $\beta $-glucosidase from Pseudomonas sp. by using a synthetic two-cistron svstem in Escherichia coli and Saccharomyces cerevisiae. Two-cistron system, 5'--tac promoter-endoglucanase gene--$\beta $-glucosidase gene-- 3', 5'-tac promoter--$\beta $-glucosidase gene--endoglucanase gene--3' and 5'-tac promoter--endoglucanase gene--SD sequence--$\beta $-glucosidase gene--3, were constructed, and expressed in E. coli and S. cerevisiae. The E. coli and S. cerevisiae contained two-cistron system produced simultaneously endoglucanase and $\beta $-glucosidase. The recombinant genes contained the bacterial signal peptide sequence produced low level of endoglucanase and $\beta $-glucosidase in S. cerevisiae transformants: Approximately above 44% of two enzymes was localized in the intracellular fraction. The production of endoglucanase and $\beta $-glucosidase in veast was not repressed in the presence of glucose or cellobiose. The veast strain contained recombinant DNA with two genes hydrolyzed carboxvmethyl cellulose, and these endoglucanase and $\beta $-glucosidase degraded CMC synergistically to glucose, cellobiose and oligosaccharide. This result suggests the possibility of the direct bioconversion of cellulose to ethanol by the recombinant yeast.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.505-510
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• 1992

An endo-$\beta$-1,4-glucanase gene of Clostridium thermocellum was cloned in Escherichia coli and was considered as a novel gene by comparison with the restriction patterns of the C. thermocellum cellulase genes so far reported. The endoglucanase from recombinant E. coli was purified by column chromatography after heat treatment. The purified enzyme was a monomer having molecular weight of 40,000. The enzyme hydrolyzed CMC to glucose and cello-oligosaccharides at :naximum activities at pH 5.0 and $65^{\circ}C$. One of the endproducts, glucose, showed no inhibitory effect on the enzyme activity, while the other endproduct, cellobiose, inhibited slightly. The values of $K_{m}$ and $V_{max}$ of the enzyme for CMC were 0.39% (w/v) and 268 Ulmg protein, respectively.