• Journal of Microbiology and Biotechnology
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• v.6 no.6
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• pp.432-438
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• 1996

Native crystalline chitin was hydrolyzed in an agitated bead reaction system using crude chitinase excreted from Aspergillus fumigatus JC-19. The reaction was enhanced significantly, and the concentration and yield of reducing sugar after 48 hours were measured to be 35.42 g/I (w/v) and 0.64, respectively, around 1.86 times higher than those of the conventional system that was carried out without glass beads. The effect of reaction conditions, such as the amounts of chitin, chitinase and glass beads, and the size of glass bead, were examined. Ball milled chitin was also hydrolyzed in the agitated bead reaction system, the conversion yield and reaction rate of ball milled chitin for 24 hours increased up to 0.87 and 48.02 g/I, respectively. Chitinase showed relatively high stability in the agitated bead reaction system, particularly in the presence of enzyme stabilizer, $Ca^{++}$, which played a critical role in preventing the deactivation of chitinase by the physical impact of glass beads. The variations of the structural features of chitin during the reaction were followed by SEM and X-ray diffraction, and the enhanced hydrolysis reaction was caused by both the fragmentation of chitin particles and the destruction of the crystalline structure owing to the synergic effects of the attrition of glass beads and the hydrolytic action of chitinase.

• Microbiology and Biotechnology Letters
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• v.18 no.3
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• pp.260-267
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• 1990

The mechanism of enzymatic hydrolysis of raw corn starch by the purified glucoamylase and a - amylase in an agitated bead reaction system was studied by investigating the changes of sugar profiles produced by each enzyme, the granular structure of raw corn starch, the amount of enzyme adsorption on residual starch, and the amylose content in residual raw starch. The sugar profiles produced by the action of exo-type glucoamylase or endo-type $\alpha$ -amylase in an agitated bead system were not recognizably differed with those produced in reaction system without bead. Without enzyme the intergenic microcrystalline structure of starch granule was not changed by the simple mechanical impact of solid media, but it was cleaved. However, starch granule was fragment into large number of small particles by the synergistic action of enzyme and attrition-milling media, identified to be the major saccharification enhancing mechanism along with the increased amount of enzyme adsorption. The amylose content decreased more readily in an agitated bead reaction system, especially by $\alpha$ -amylase.

• Microbiology and Biotechnology Letters
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• v.19 no.2
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• pp.163-170
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• 1991

Production of cyclodextrin (CD) directly from raw corn starch without liquefaction using cyclodextrin glucanotransferase (CGTase) was carried out in an agitated bead reaction system. Similar CD yield and production rate comparable with those of conventional method using liquefied starch were obtained. Especially high purity-CD in the reaction mixture without accumulation of malto-oligosaccharides was obtained. The maximum 54g/l of CD was obtained at raw starch concentration of 200g/l. CD yield was inversely proportional to raw starch concentration, and conversion yield was 0.48 at substrate concentration of 100g/l. The optimal amount of enzyme (CGTase unit/g raw starch) was found to be around 6.0. Granular structure of raw starch degraded by CGTase was observed by SEM in order to investigate the enhancing mechanism, along with those of acid or alkali pretreated raw starch, amylose, and amylopectin. Kinetic constants of CGTase on raw starch in an agitated bead reaction system were evaluated, and CGTase was competitively inhibited by CD.

• Microbiology and Biotechnology Letters
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• v.14 no.5
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• pp.407-413
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• 1986

In an agitated bead reaction system, the enzymatic saccharification of uncooked starch was substantially enhanced. The enhancement mechanism was investigated front the view of the structural aspect of starch. The mechanical impact caused by the movement of the attrition-milling media resulted neither the destruction of microcrystalline structure nor the fragmentation of starch granule. instead, the most distinct phenomenon was the swelling of starch granule up to about 2.5 times, and the swelling mechanism was not similar with that caused by cooking. However, in the case of the enzyme addition in the attrition coupled reaction system, the swollen starch was easily fragmented into the large number of small particles by the synergistic action of the enzyme and milling-media. The exposed surface area of the fragmented particles plays the major role in enhancing the saccharification. The saccharification rate was quite different depending on the source of starch, the reason was discussed in terms of the granular structure of uncooked starches.

• Microbiology and Biotechnology Letters
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• v.18 no.4
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• pp.352-359
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• 1990

The synergistic effect of glucoamylase and a -amylase on the hydrolysis of raw corn starch in an agitated bead reaction system was studied by investigating the changes of sugar profiles, the granular structure, particle size distribution, and X-ray diffraction pattern of residual raw corn starch. The enzymatic hydrolysis of raw corn starch was greatly enhanced by synergistic effect of glucoamylase and $\alpha$ -amylase. Even though the sugar profiles were mainly determined by the mixing ratio of glucoamylase and $\alpha$-amylase; raw starch was mainly converted to glucose directly without accumulation of any significant amount of oligosaccharides. The cavity formation and fragmentation phenomena of raw corn starch granule subjected to enzyme reaction were analyzed by means of SEM and the particle size distribution. The X-ray diffraction pattern of raw starch was not changed at the initial stage of reaction but slightly changed at the late stage of hydrolysis, which may be caused by the preferential degradation of amorphous region by enzymatic reaction, not by the destruction of microcrystalline structure of raw corn starch.

• Microbiology and Biotechnology Letters
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• v.14 no.5
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• pp.399-405
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• 1986

Corn starch was saccharified without cooking in an agitated bead reaction system. Uncooked corn starch was effectively hydrolyzed even at the concentration as high as 39%(w/v). After 24 hours. the extent of saccharification reached at 92%, which corresponds glucose concentration of 425g/L. Fed-batch feeding of starch was more effective than batch feeding for saccharification of uncooked corn starch. The composition of hydrolysated of uncooked starch was analyzed. which was composed of 95% glucose, 0.7% of maltose, and 4.5% of high saccharide, similar with that of cooked starch. The hydrolysate can be successfully utilized for HFCS manufacture. The starch liquefying and saccharifying enzyme was relatively stable even be the physical impact of the attrition-milling media. The enzyme stabilizer, $Ca^{++}$, played an essential role in preventing the enzyme deactivation caused by the physical impact.

• KSBB Journal
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• v.4 no.2
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• pp.78-86
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• 1989

The effective saccharification of cellulosic biomass to glucose is the most critical step for the conversion of renwable biomass to alternative liquid fuel. The enzymatic hydrolysis of biomass can be significantly enhanced provide the attrition milling media is added during hydrolysis. The enhancing mechanism of hydrolysis reaction in an agitated bead system was investigated. An attrition-reactor (bioattritor) which installed specially designed torque measuring apparatus was developed, and the potimal saccharification conditions of bioattritor were determined. The relationship between the power consumption required for agitation of attrition-milling media and enhanced extent of hydrolysis of biomass was compared to evaluatic economic feasibility of the process.

• Microbiology and Biotechnology Letters
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• v.17 no.4
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• pp.349-357
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• 1989

Uncooked starch can be effectively saccharified in an enzyme reaction system containing attrition-milling media. To develope the high efficiency bioattritor, an agitated bead type bioreactor was constructed, and its effectiveness was evaluated. The optimal operation condition of bioattritor was found to be 300 g glass bead/L, 200 rpm, standard type impeller for 220 g/L of uncooked corn starch. The torque under the various operational conditions were also measured. The interrelation-ship between energy consumption for agitation of attrition-milling media and enhanced extent of saccharification of uncooked starch was evaluated, Power consumption was measured to be around 1.53 watt/L under the optimal operation condition. The attrition coupled enzyme reaction system is identified to tie a very excellent energy saying process for saccharification of uncooked starch, and seems to have a bright prospect of industrial application.