• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.704-708
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• 1999

In addition to catalyzing the synthesis of glucan from sucrose as a primary reaction, glucansucrase also catalyzes the transfer of glucose from sucrose to other carbohydrates that are present or are added to the reaction digest. Using dextransucrase and altemansucrase, prepared from Leuconostoc mesenteroides B-742CBM and B-1355C, respectively, we modified the bacterial cellulose in Acetobacter xylinum ATCC10821 culture, and then produced a characteristic cellulose that is soluble and has a new structure. There were also some partially modified insoluble cellulose and oligosaccharides in the modification culture. After methylation and following acid hydrolysis of both the soluble and insoluble glucans, there were ($1{\rightarrow}4$) as well as ($1{\rightarrow}6$) and ($1{\rightarrow}3$) glycosidic linkages in the soluble glucan.

• Journal of Microbiology and Biotechnology
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• v.9 no.5
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• pp.529-533
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• 1999

In addition to catalyzing the synthesis of dextran from sucrose as a primary reaction, dextransucrase also catalyzes the transfer of glucose from sucrose to other carbohydrates that are present or are added to the reaction digest. We have synthesized new glucans having new structures and new characteristics, by transferring D-glucose of sucrose to $\alpha$-cellulose and by using the constitutive dextransucrase obtained from Leuconostoc mesenteroides B-742CBM. The final reaction products were composed of soluble- and insoluble-glucans. The yields of soluble- and insoluble-glucans were theoretically 21% $\pm$ 2.2 and 68% $\pm$ 5.1, respectively. The remainder of the reaction products was recovered as a mixture of olgiosaccharides that could not be precipitated by 67%(v/v) ethanol. Treating the modified glucans with endo-dextranase and/or cellulase, oligosaccharides were produced that were not formed from the hydrolysis of native cellulose or B-742CBM dextran. The modification of the cellulose was confirmed by methylation and acid hydrolysis of the soluble-and insoluble-glucan. Both (1->4) and(1->6) glycosidic linkages were found in both of the glucans.

• Preventive Nutrition and Food Science
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• v.1 no.1
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• pp.117-120
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• 1996

To enhance cellulose hydrolysis f Humicola insolens cellulase (HIC) was empolyed with immobilized glucose isomerase(IGI). Optimun pH and temperature for HIC were 6.5 and 55$^{\circ}C$, respectively, whereas those for IGI were 7.0 and 60$^{\circ}C$, respectively. Optimun loading size of IGI was 200mg(130 untis) with 15units of HIC. Reaction conditions were determined to be as follows: 55$^{\circ}C$,pH 6.5, HIC 15 units and IGI 130 units. After 24h hydrolysis, more than 65% of avicel was converted to glucose and fructose; in contrast, the conversion ratio of control was 40%.

• Microbiology and Biotechnology Letters
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• v.8 no.1
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• pp.41-46
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• 1980

Low molecular weight endo-glucanase was partially purified from cellulase complex using Sephadex G-100 gel chromatography. Biochemical properties of the purified component was investigated. Optimum pH and temperature determined were 6.0 and 5$0^{\circ}C$, respectively. Enzymatic hydrolysis of four cellulosic substrates having varying crystallinity was evaluated. It was found that hydrolysis of amorphous region was followed by the hydrolysis of crystalline region. In order to examine the effect of adsorption of the enzyme onto the cellulosic substrates on the hydrolysis kinetics, adsorption studies were carried out. Time course of adsorption of low molecular weight endo-glucanase onto various cellulostances was observed for 25 min. The rate and amount of adsorption to amorphous cellulose was greater than those to the crystalline cellulose. This result suggested that the role of endo-glucanasc was more important to the hydrolysis of amorphous cellulose than to the crystalline region of the cellulose.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.4
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• pp.329-333
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• 2005

This study was designed to investigate the stability of a lipase fused with a cellulose­binding domain (CBD) to cellulase. The fusion protein was derived from a gene cluster of a CBD fragment of a cellulase gene in Trichoderma hazianum and a lipase gene in Bacillus stearother­mophilus L1. Due to the CBD, this lipase can be immobilized to a cellulose material. Factors affecting the lipase stability were divided into the reaction-independent factors (RIF), and the re­action-dependent factors (RDF). RIF includes the reaction conditions such as pH and tempera­ture, whereas substrate limitation and product inhibition are examples of RDF. As pH 10 and $50^{\circ}C$ were found to be optimum reaction conditions for oil hydrolysis by this lipase, the stability of the free and the immobilized lipase was studied under these conditions. Avicel (microcrystal­line cellulose) was used as a support for lipase immobilization. The effects of both RIF and RDF on the enzyme activity were less for the immobilized lipase than for the free lipase. Due to the irreversible binding of CBD to Avicel and the high stability of the immobilized lipase, the enzyme activity after five times of use was over $70\%$ of the initial activity.

• BMB Reports
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• v.31 no.1
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• pp.53-57
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• 1998

The CMCax gene from Acetobacter xylinum ATCC 23769 was cloned and expressed in E. coli. With this gene, three gene products - mature CMCax, CMCax containing signal peptide(pre-CMCax), and a glutathione-S-transferase(GST)-CMCax fusion enzyme - were expressed. CMCax and pre-CMCax are aggregated to multimeric forms which showed high CMC hydrolysis activity, whereas GST-CMCax was less aggregated and showed lower activity, indicating that oligomerization of CMCax controbutes to the cellulose hydrolysis activity to achieve greater efficiency. The enzyme was identified to be an $\beta$-1,4-endoglucanase, which catalyzes the cleavage of internal $\beta$-1,4-glycosidic bonds of cellulose. The reaction products, cellobiose and cellotriose, from cellopentaose as a substrate, were identified by HPLC. Substrate specificity of cellotetraose by this enzyme was poor, and the reaction products consisted of glucose, cellobiose, and cellotriose in a very low yield. Theses results suggested that cellopentaose might be the oligosaccharide substrate consisting of the lowest number of glucose. The optimum pH of CMCax and pre CMCax was about 4.5, whereas that of GST-CMCas was rather broad at pH 4.5-8. The physiological significance of cellulose-hydrolyzing enzyme, CMCax, having such low $\beta$-1,4-endoglucanase activity and low optimum pH in cellulose-producing A. xylinum is not clearly known yet, but it seems to be closely related to the production of cellulose.

• KSBB Journal
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• v.4 no.3
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• pp.221-228
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• 1989

Celluose is an insoluble substrate, therefore, a proper mixing of the cellulose suspension is essential for an effective enzymatic hydrolysis. To study the effect of mixing motion of various enzyme reactors on enzymatic hydrolysis of cellulose, three distinct types of biroreator: vertical impeller type bioreator(VITB), horizontal paddle type bioreactor(HPTB), and tumbling drum type bioreactor(TDTB), were assembled and their performance was compared. The optimal agitation speed was 100rpm for VITB and HPTB, 200rpm for TDTB. The saccharification efficiency of each reactor was compared under the optimal agitation intensity. The highest degree of saccharification was achieved in the case of VITB, especially, at high cellulose concentration. The VITB seems to be the most suitable type of bioreactor that can maintain proper mixing pattern for effective enzyme reaction. In the view of energy consumption, the TDTB showed the lowest value: however, the energy consumption was rapidly increased at high concentration of celluose. To dertermine the most suitable type of bioreactor, the entire process, including substrate cost, substrate concentration, and feasibility of scale-up, needs to be evaluated.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1979.10a
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• pp.243.3-244
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• 1979

Low molecular weight endo-glucanase fraction of cellulase from Trichoderma reesei was purified using Sephadex G-100 and concanavalin A-Sepha-rose 4B affinity chromatography. Its biochemical characteristics including pH profile, temperature profile and kinetic behavior were studied. The optimum conditions for enzymatic reaction were pH 6.0 and $5^{\circ}C.$ The activation energy for CMC (carboxymethylcellulose) was 10,800 cal/mole. Its adsorption to amorhous and crystalline cellulose was observed. Adsorption to amorphous cellulose was more rapid and greater than that of crystalline cellulose. Reconstitution study was performed. Significance of low molecular weight endo-glucanase on cellulose hydrolysis will be further discussed.

• Journal of Microbiology and Biotechnology
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• v.9 no.1
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• pp.1-8
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• 1999

Most carbohydrates exist in nature in an insoluble state, which reduces their susceptibility towards various carbohydrases. Accordingly, they require intensive pretreatment for structural modification to enhance an enzyme reaction. The direct conversion of insoluble carbohydrates has distinct advantages for special types of reaction, especially exo-type carbohydrase; however, its application is limited due to structural constraints. This paper introduces two novel heterogeneous enzyme reaction systems for direct conversion of insoluble carbohydrates; one is an attrition coupled enzyme reaction system containing attrition-milling media for enhancing the enzyme reaction, and the other is a heterogeneous enzyme reaction system using extruded starch as an insoluble substrate. The direct conversion of typically insoluble carbohydrates, including cellulose, starch, and chitin with their corresponding carbohydrases, including cellulase, amylase, chitinase, and cyclodextrin glucanotransferase, was carried out using two proposed enzyme reaction systems. The conceptual features of the systems, their reaction characteristics and mechanism, and the industrial applications of the various carbohydrates are analyzed in this review.

• Journal of the Korean Wood Science and Technology
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• v.40 no.2
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• pp.78-90
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• 2012

Soybean hull is an attractive feedstock for glucose production. To increase the glucose conversion in acid hydrolysis, a pretreatment method combined steam explosion with alkali pretreatment for soybean hull was studied. For first step pretreatment, steam explosion conditions (log Ro 2.45) were optimized to obtain maximum solid recovery and cellulose content. In the second step pretreatment, the conditions for potassium hydroxide pretreatment of steam exploded soybean hull were optimized by using RSM (response surface methodology). The optimum conditions for minimum lignin content were determined to be 0.6% potassium hydroxide concentration, $70^{\circ}C$ reaction temperature and 198 min reaction time. The predicted lignin content was 2.2% at the optimum conditions. Experimental verification of the optimum conditions gave the lignin content in similar value with the estimated value of the model. Finally, glucose conversion of pretreated soybean hull using acid hydrolysis resulted in $97.1{\pm}0.4%$. This research of two-step pretreatment was a promising method for increasing the glucose conversion in the cellulose-to-glucose process.