• Journal of Microbiology and Biotechnology
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• v.29 no.4
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• pp.625-632
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• 2019

The unified saccharification and fermentation (USF) system was developed for direct production of ethanol from agarose. This system contains an enzymatic saccharification process that uses three types of agarases and a fermentation process by recombinant yeast. The $pGMF{\alpha}-HGN$ plasmid harboring AGAH71 and AGAG1 genes encoding ${\beta}-agarase$ and the NABH558 gene encoding neoagarobiose hydrolase was constructed and transformed into the Saccharomyces cerevisiae 2805 strain. Three secretory agarases were produced by introducing an S. cerevisiae signal sequence, and they efficiently degraded agarose to galactose, 3,6-anhydro-L-galactose (AHG), neoagarobiose, and neoagarohexose. To directly produce ethanol from agarose, the S. cerevisiae $2805/pGMF{\alpha}-HGN$ strain was cultivated into YP-containing agarose medium at $40^{\circ}C$ for 48 h (for saccharification) and then $30^{\circ}C$ for 72 h (for fermentation). During the united cultivation process for 120 h, a maximum of 1.97 g/l ethanol from 10 g/l agarose was produced. This is the first report on a single process containing enzymatic saccharification and fermentation for direct production of ethanol without chemical liquefaction (pretreatment) of agarose.

• Microbiology and Biotechnology Letters
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• v.20 no.4
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• pp.437-444
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• 1992

An innovative process for high fructose corn syrup (HFCS) production coupled with direct saccharification of raw corn starch in the agitated bead enzyme reactor (bioattitor) was investigated. The required high concentration/purity of glucose solution suitable for isomerization was produced directly in a bioattritor. without condensation of hydrolyzate, 398 g glucose/$\ell$ and 98% glucose content from 400 g/$\ell$ (w/v) of raw corn starch after 24 hours. The unsaccharified residual starch could be separated easily upon centrifugation, and resaccharified. The obtained solution also possessed other desirable requirements as substrate for isomerization, such as. low concentrations of denatured protein and calcium ions, thereby, simplified the purification step. The obtained glucose solution was isomerized in an enzyme reactor paked with immobilized glucose isomerase to evaluate the suitability as a substrate. The proposed new HFCS process seems to have many advantages over the conventional process via liquefaction-saccharification steps. The follow-up investigations of the proposed process need to be conducted to evaluate the feasibility of industrial application.

• Microbiology and Biotechnology Letters
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• v.48 no.1
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• pp.32-37
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• 2020

We improved on a unified saccharification and fermentation (USF) system for the direct production of ethanol from agarose by increasing total agarase activity. The pGMFα-NGH plasmid harboring the NABH558 gene encoding neoagarobiose hydrolase and the AGAG1 and AGAH71 genes encoding β-agarase was constructed and used to transform Saccharomyces cerevisiae 2805. NABH558 gene transcription level was increased and total agarase activity was increased by 25 to 40% by placing the NABH558 gene expression cassette upstream of the other gene expression cassettes. In the 2805/pGMFα-NGH transformant, three secretory agarases were produced that efficiently degraded agarose to galactose, 3,6-anhydro-L-galactose (AHG), neoagarobiose, and neoagarohexaose. During the united cultivation process, a maximum of 2.36 g/l ethanol from 10 g/l agarose was produced over 120 h.

• Microbiology and Biotechnology Letters
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• v.21 no.2
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• pp.157-162
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• 1993

Raw starch can be effectively saccharified in an enzyme reaction system containing sttrition-milling media. In order to develop an effcient attrition-coupled bioreactor(bioattritor), a rotating drum type bioattitor was construced, and its optimal operation conditions and power consumptions were evaluated. The optimal conditions for 3l bioattritor were 4 baffled, baffle size of 1:0.05 (the ratio of drum diameter to baffle), drum rotation speed of 100 rpm, and 1.33g of 3 mm glass bead/g of raw corn starch.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.1-8
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• 2017

The production of high-value chemicals from natural resources as an alternative for petroleum-based products is currently expanding in parallel with biorefinery. The use of lignocellulosic biomass as raw material is promising to achieve economic and environmental sustainability. Filamentous fungi, particularly Aspergillus species, are already used industrially to produce organic acid as well as many enzymes. The production of lignocellulose-degrading enzymes opens the possibility for direct fungal fermentation towards organic acids such as itaconic acid (IA) and fumaric acid (FA). These acids have wide-range applications and potentially addressable markets as platform chemicals. However, current technologies for the production of these compounds are mostly based on submerged fermentation. This work showed the capacity of two Aspergillus species (A. terreus and A. oryzae) to yield both acids by solid-state fermentation and simultaneous saccharification and fermentation. FA was optimally produced at by A. oryzae in simultaneous saccharification and fermentation (0.54 mg/g wheat bran). The yield of 0.11 mg IA/g biomass by A. oryzae is the highest reported in the literature for simultaneous solid-state fermentation without sugar supplements.

• Journal of the Korean Wood Science and Technology
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• v.32 no.1
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• pp.28-34
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• 2004

This study was performed to evaluate the characteristics of the carboxymethylated substrate from high reactive autohydrolyzed cellulose (HRC) and those of commercial α-cellulose (CAC) and refiner mechanical pulp (RMP). Saccharification rates of HRC substrate were achieved over 70% with 12 hr hydrolysis, about 90% with 24 hr, and 99.5% with 72 hr. CMCase and avicelase activities of cellulase onozuka were 4.09 ㎛ G/mg·min and 14.0 ㎛ G/mg·min, respectively. There were no any significant changes in cellulase activities with this substrate. The saccharification rates of CAC and RMP were very low, 57% and 38% with 72 hr, respectively. Those lignin-zero autohydrolyzed substrates, HRC and CAC, were highly carboxymethylated at the high alkali concentration, near 30%, for 3 hr. reaction, and resulted in 1.13-1.15 of D.S., besides 0.85 of D.S. from RMP. Water solubilities of carboxymethylated substrates were increased with an increase of D.S., 98-98.5% from HRC and CAC and 31.5% from RMP. RMP which has low specific surface area showed lower water retention values, compared to high values of 435 and 321% from CAC and HRC, respectively. There were no direct relationship between surface area and swelling ratio of the substrates.