• Journal of Microbiology and Biotechnology
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• 제26권7호
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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.

• Journal of Microbiology and Biotechnology
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• 제2권3호
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• pp.197-203
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• 1992

Studies of the pretreatment of chitin and its subsequent hydrolysis by Aspergillus carneus chitinase are reported. Ball milling was found to be the most effective way among the pretreatment methods tested. Data are presented describing the effect of enzyme and substrate concentrations on the rate and extent of the hydrolysis process. It was found that the successive addition of enzyme improved the saccharification yield. Significant product inhibition of the chitinase was observed when N-acetylglucosamine concentration was 3.6％ or higher. Adsorption of enzymes to the substrate occurred during a 24 hr hydrolysis period. An initial rapid and extensive adsorption occurred, followed by gradual desorption which increased during the time of reaction. Intermediate removal of the hydrolyzate and continuation of the hydrolysis by adsorbed enzyme on the residual chitin was also investigated. A total of 75.4 g/l reducing sugars, corresponding to 69.2％ saccharificaton yield (as N-acetylglucosamine) was obtained. In addition an increase in the amount of recoverable enzymes was observed under these conditions. Evidence presented here suggests that the technique, whereby the free enzymes in the recovered hydrolyzate are re-adsorbed onto the new substrate, may provide a means of recirculating the dissolved enzymes.

• Journal of Forest and Environmental Science
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• 제36권1호
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• pp.62-66
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• 2020

Ethanol fermentation of the enzymatic hydrolysates from the products pretreated using 1-ethyl-3-methyl-imidazolium acetate ([EMIM]Ac) and its co-solvents with dimethylformamide (DMF) was conducted using Saccharomyces cerevisiae (D452-2). The optical density change due to the yeast cell growth, the consumption amount of monosugars (glucose, xylose), the concentration of acetate, and ethanol production yield were investigated. The co-solvent system lowered inhibition of the growth of the cells. The highest concentration of glucose (7.8 g/L) and xylose (3.6 g/L) was obtained from the enzymatic hydrolysates of the pretreated product by pure [EMIM]Ac. The initial concentration of both monosugars in the enzymatic hydrolysates was decreased with increasing fermentation time. Ethanol of Approximately 3 g/L was produced from the enzymatic hydrolysates by pure [EMIM]Ac and co-solvent with less than 50% DMF.

• 한국미생물·생명공학회지
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• 제17권4호
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• pp.349-357
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• 1989

전분질을 당화시킬 때 유리구와 같은 분쇄마찰매체 (attrition-milling media)를 첨가하여 교반함으로써 생전분질 -효소 현탄액에 분쇄마찰효과를 주어 전분입자를 구조적으로 변화시켜 당화를 현저히 촉진시키는 적절한 무증자 당화법을 연구해오고 있다. 본 연구는 전분질의 무증자 당화에 적합한 고효율 및 저에너지 소모형 분쇄마찰매체 함유 효소반응기의 개발을 목포로, impeller를 갖춘 agitated bead type bloattritor를 설계 제작하여 그 효용성을 검토하였다. 무증자 옥수수전분을 기질로 당화에 적합한 bioattritor의 최적 조작조건을 검토하였고, 일정한 탄성계수를 지닌 spiral spring coil을 내장한 torque 측정장치를 개발하여 부하되는 torque 및 분쇄마찰 매체의 교반에 소요되는 energy를 측정하였고 중요 변수의 영향을 검토하였다. 또한 energy 소모와 생 전분의 효소당화 촉진효과와의 상관관계를 규명하였으며, bioattritor의 경제성을 전망하였다. 전분질의 최적 당화조건에서 분쇄마찰매체의 교반에 소요되는 동력은 1.53watt/ι로서 매우 적었다. 분쇄마찰매체 함유 효소반응계는 에너지 절약형 공정으로 사료되며, 이를 이용한 무증자 전분 효소당화공정의 산업적 활용 전망이 예상된다.

• KSBB Journal
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• 제4권3호
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• pp.215-220
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• 1989

불용성 기질인 섬유소의 고효율 효소당화에 적합한 저에너지 소모형 분쇄마찰매체 함유 효소반응기의 개발을 목표로 수평형 회전축에 부착된 paddle에 의해 반응조 내부의 섬유소-분쇄마찰매체-효소의 혼합액을 교반 마찰시키는 horizontal paddle type bioreactor를 제작하여 효용성과 당화조건을 검토하였다. 여러 실험조건하에서 당생성효율과 분쇄마찰매체의 교반시 소모되는 에너지를 다른 두 형태의 bioreactor와 비교 분석함으로서 공업적 활용을 위한 기술적, 경제적 타당성을 비교평가하였다. 표준당화조건은 3mm의 분쇄마찰매체를 500g / L로 첨가하여 200rpm에서 교반할 때였으며, 특히 HPTB는 고농도 기질의 당화에 적합한 bioreactor로 판단된다. 표준당화조건에서 분쇄마찰매체의 교반시 소모되는 동력은 1.7watch/g였으며, 고농도 기질을 이용할 경우 더욱 낮은 동력소모로 당을 생산할 수 있었다. 뿐만 아니라 scale up이 용이할 것으로 예상되는 등 섬유소의 고농도 당화에 적합한 고효율 분쇄마찰매체함유 효소반응기로 판단된다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권1호
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• pp.26-31
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• 2006

The saccharogenic liquid (SFW) obtained by the enzymatic saccharification of food wastes was used as a medium for production of bacterial cellulose (BC). The enzymatic saccharification of food wastes was carried out by the cultivation supernatant of Trichoderma harziaum FJ1 culture. Acetobacter xylinum KJ1 was employed for the BC production culture. The physical properties, such as polymerization, crystallinity, Young's modulus, and tensile strength, of BCs produced by three culture methods: the static cultures using HS (Hestrin-Schramm) as a reference medium (A) or the SFW medium (B), the shaking culture (C) or the air circulation culture (D) using the SFW medium, were investigated. The degrees of polymerization of BCs produced under the different culture conditions (A-D) showed 11000, 9500, 8500, and 9200, respectively. Young's modulus was 4.15, 5.0, 4.0, and 4.6 GPa, respectively. Tensile strength was 124, 200, 80, and 184 MPa, respectively. All of the BC had a form of cellulose I representing pure cellulose. In the case of the shaking culture, the degree of crystallinity was 51.2%, the lowest degree. Under the other culturing conditions, the trend should remain in the range of 89.7-84%. Overall, the physical properties of BC produced from SFW were similar to those of BC from HS medium, a commercial complex medium, and BC production by the air circulation culture mode brought more favorable results in terms of the physical properties and its ease of scale-up. Therefore, it is expected that a new BC production method, like air circulation culture using SFW, would contribute greatly to BC-related manufacturing.

• KSBB Journal
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• 제11권6호
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• pp.712-717
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• 1996

셀룰라아제, polyoxyethylene 유도체와 maleic acid anhydride의 공중합고분자로 중합한 수식셀룰 라아제를 제조하고, 당화 특성과 효소 반응 속도론 등을 검토하였다. 수식 반응에서, 공중합고분자-효소 질량비가 증가함에 따라 수식률은 증가하며, 질량비 4 이상에서는 수식률이 일정하였다. 이 때 최 대수식률은 55% 이며 최대수식률의 수식생룰라아제 는 75%의 높은 효소활성을 나타내었다. 수식효소와 미수식효소의 당화 실험에서, 수식효소가 전 반응시 간에 걸쳐 미수식효소보다 높은 당화반응안정성을 유지하였으며, 그로 인해 최종 전화율은 수식효소가 더 큰 값을 가졌다. 기질에의 강한 흡착으로 인한 효소 deactivation을 고려한 반응식을 적용한 결과, 생 성환원탕놓도의 실험값과 계산치는 잘 일치하였다. 그로부터 반응속도상수를 구하고 생성환원당농도와 유리효소농도의 모사그래프를 구할 수 있었으며, 반 응시간에 따른 유리효소의 농도를 수치모사한 결과, 미수식효소에 비해 수식효소의 유리효소 농도가 더 높았고 그로 인해 더 높은 당전화율을 나타내였다.

• KSBB Journal
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• 제10권3호
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• pp.304-316
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• 1995

액화 실험에서 BAN과 Termamyl을 비교할 때 전반적으로 Termamyl이 우수했으며 Termamyl을 (0.00625% ~0.025% 범위로 첨가하였을 때 첨가량 증가에 따른 액화반응 촉진이 뚜렷이 관찰되였다. Termamyl 240uc의 경우 타파오카에 대해 최소 0 0.0125 (v/w)% 이상이 필요하였고, 반응시간은 2 시간 정도가 석당하였다. 최적온도는 $90∼95^{\circ}C$이었고 pH는 5.8 부근이 최척이였다. 반응초기 점도가 매우 높게 나타나는데 호화속도를 약간 줄이가 위해 초기 온도를 $70^{\circ}C$정도로 낮추는 갯이 좋았다. 당화실험에서는 Novo AMG 및 국내 도일 산엽 당 화효소 모두 액화된 타피오카 용액을 잘 당화시켰다. 그러나 Novo AMG와 도일 당화효소의 활성은 기 질 에 따라 약간썩 탈랐다. 즉 이당류인 말토오스를 기 질 로 할 때는 Novo AMG의 단위 부피당 활성이 도일 당화효소의 약 1.2배이였으나, 전분을 기질로 할 때는 활성비가 1.5배였다. 경시적으로 볼 때 초기에는 당화 속도가 빨랐고 포도망이 축적되는 후기에는 당화속도 가 감소하였다. 당화의 최적온도는 약 $55∼60^{\circ}C$ 부근이었고 pH는 조절하지 않는 경우 (pH 5.7)와 조절 한 경우 (pH 4.3)간에 차이가 없었다. 85% 이상의 당화를 위해서는 타피오카 샤용량에 대해 Novo AMG 400uc을 기준으로 O.0625(v/w)% 이상, 그 리고 10시간 이상의 반응사간이 필요하였다.

• Journal of Microbiology and Biotechnology
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• 제28권10호
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• pp.1671-1682
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• 2018

Alginate lyases (endo and exo-lyases) are required for the degradation of alginate into its constituting monomers. Efficient bioethanol production and extraction of bioactives from brown algae requires intensive use of these enzymes. Nonetheless, there are few commercial alginate lyase preparations, and their costs make them unsuitable for large scale experiments. A recombinant expression protocol has been developed in this study for producing seven endo-lyases and three exo-lyases as soluble and highly active preparations. Saccharification of alginate using 21 different endo/exo-lyase combinations shows that there is complementary enzymatic activity between some of the endo/exo pairs. This is probably due to favorable matching of their substrate biases for the different glycosidic bonds in the alginate molecule. Therefore, selection of enzymes for the best saccharification results for a given biomass should be based on screens comprising both types of lyases. Additionally, different incubation temperatures, enzyme load ratios, and enzyme loading strategies were assessed using the best four enzyme combinations for treating Macrocystis pyrifera biomass. It was shown that $30^{\circ}C$ with a 1:3 endo/exo loading ratio was suitable for all four combinations. Moreover, simultaneous loading of endo-and exo-lyases at the beginning of the reaction allowed maximum alginate saccharification in half the time than when the exo-lyases were added sequentially.

• Journal of Microbiology and Biotechnology
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• 제21권7호
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• pp.703-710
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• 2011

Enzymatic saccharification of corn stover using Phanerochaete chrysosporium and Gloeophyllum trabeum and subsequent fermentation of the saccharification products to ethanol by Saccharomyces cerevisiae and Escherichia coli K011 were achieved. Prior to simultaneous saccharification and fermentation (SSF) for ethanol production, solid-state fermentation was performed for four days on ground corn stover using either P. chrysosporium or G. trabeum to induce in situ cellulase production. During SSF with S. cerevisiae or E. coli, ethanol production was the highest on day 4 for all samples. For corn stover treated with P. chrysosporium, the conversion to ethanol was 2.29 g/100 g corn stover with S. cerevisiae as the fermenting organism, whereas for the sample inoculated with E. coli K011, the ethanol production was 4.14 g/100 g corn stover. Corn stover treated with G. trabeum showed a conversion 1.90 and 4.79 g/100 g corn stover with S. cerevisiae and E. coli K011 as the fermenting organisms, respectively. Other fermentation co-products, such as acetic acid and lactic acid, were also monitored. Acetic acid production ranged between 0.45 and 0.78 g/100 g corn stover, while no lactic acid production was detected throughout the 5 days of SSF. The results of our experiment suggest that it is possible to perform SSF of corn stover using P. chrysosporium, G. trabeum, S. cerevisiae and E. coli K011 for the production of fuel ethanol.