• Title/Summary/Keyword: Saccharification

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Improvement of Cellobiose Dehydrogenase(CDH) and $\beta$-Glucosidase Activity by Phanerochaete chrysosporium Mutant (Phanerochaete chrysosporium 변이주에서의 Cellobiose Dehydrogenase(CDH)와 $\beta$-Glucosidase 활성 향상)

  • Kim, Eun-Ji;Kang, Seong-Woo;Song, Kwang-Ho;Han, Sung-Ok;Kim, Jae-Jin;Kim, Seung-Wook
    • Korean Chemical Engineering Research
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    • v.49 no.1
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    • pp.101-104
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    • 2011
  • Cellobiose dehydrogenase(CDH) as a hemoflavoenzyme is secreted out of cell in the cellulose degradation. As CDH strongly bound to amorphous cellulose, it helps cellulose hydrolysis by cellulase. CDH may have an important role of saccharification process for bioethanol production. In this study, Phanerochaete chrysosporium ATCC 32629 was selected for the production of CDH among other strains tested. The optimal temperature and pH of CDH produced by P. chrysosporium ATCC 32629 were ${55^{\circ}C}$ and 4, respectively. To improve the activity of CDH, the mutation of P. chrysosporium was performed using proton beam that has high energy level partially. As a result, P. chrysosporium mutant with the high activity was selected at 1.2 kGy in a range of 99.9% lethal rate. The CDH and $\beta$-glucosidase activities of mutant were 1.4 fold and 20 fold higher than those of wild strain. Therefore, P. chrysosporium mutant with the high activities of CDH and $\beta$-glucosidase was obtained from mutation by proton beam irradiation.

Fermentation and Quality Characteristics of Korean Traditional Cheongju by Different Mashing Methods (청주 제조시 담금방법에 따른 발효 및 품질 특성)

  • Bae, Gyun-Ho;Lee, Sang-Hyeon;Cheong, Chul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.17 no.8
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    • pp.637-645
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    • 2016
  • This study was conducted to investigate the characteristics of fermentation and quality of Cheongju prepared by mashing using rice Nuruk inoculated with Aspergillus oryzae. Mashes were prepared by fermentation for 30-50 days using different amounts of fermenting agent, brewing water, milling ratios and fermenting temperatures. Adding fermenting agent at 15% resulted in slow fermentation, but a final alcohol content of 17% (v/v), similar to other samples tested. Addition of higher amounts of Nuruk resulted in increased amounts of citric acid, tartaric acid and malic acid, but low levels of succinic acid. Incomplete fermentation occurred when the ratio of brewing water was low, but the alcohol content (17%) of all samples was similar. When the amount of brewing water was high, the organic acid was levels were high. The speed of saccharification and fermentation was low when fermentation was conducted at $10^{\circ}C$, but the final alcohol content was the highest at this temperature. However, the content of n-propanol, isobutanol, isoamyl alcohol and organic acid was low at low temperature. At this time, the content of citric acid and malic acid was low, but the content of succinic acid was high. A higher milling degree resulted in a lower content of alcohol, organic acid and higher alcohols, with 10% milling resulting in a significantly higher content than the other samples.

Immobilization of an Enzyme with Chitosan Microbeads (Chitosan Microbeads에 의한 효소고정화)

  • SOHN Heung-Sik;PARK Seong-Min;SON Byung-Yil;CHOI Hyeon-Mee;LEE Keun-Tai
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.1
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    • pp.83-87
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    • 1999
  • Immobilization of amyloglucosidase (AMG) with chitosan microbead and its possible applications were evaluated. The diameter of chitosan inicrobead was about 1.2 mm and the optimum enzyme concentration for immobilization was 6 mg/ml. The relative activity of the immobilized enzyme was $97.8\%$ at pH 4.2 and $55^{\circ}C$ and the optimum condition for the immobilized enwme was the same to that of free enzyme. In case of temperature above $30^{\circ}C$, the activity of the immobilized enzyme was a little higher than that of free enzyme. The enzyme activities of both free and immobilized were stable for 6 months when stored at $35^{\circ}C$. The optimum temperatures of both enzymes for saccharification of the dextrinized starch were $55^{\circ}C$ while the relative activity of the immobilized enzlme was $62.6\%$.

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Volatile Aromatic Compounds and Fermentation Properties of Fermented Milk with Buckwheat (메밀을 첨가한 발효유의 향기성분과 발효특성에 관한 연구)

  • Lee, Beom-Seon;Park, Seung-Kook
    • Korean Journal of Food Science and Technology
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    • v.45 no.3
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    • pp.267-273
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    • 2013
  • We aimed to improve the flavor quality of plain yogurt, which is known to be sour and less desirable in flavor, varying concentrations of a buckwheat saccharification solution (BSS) were added to milk, followed by fermentation with commercially available mixed strains of lactic acid bacteria. Volatile compounds were analyzed using the gas chromatography-headspace-solid phase microextraction (GC-HS-SPME) method. Fermentation properties, including pH, titratable acidity, viable cells, viscosity, and color value were also measured. Eleven volatile compounds were identified with GC-MS. Of which, diacetyl, butanoic acid, and 2-heptanone proportionally increased as the levels of BSS increased. Undesirable compounds such as acetic acid and 2-butanone, decreased as BSS concentration increased. Fermentation properties were significantly altered with the addition of BSS. Our findings indicate that the flavor quality of plain yogurt can be improved by adding BSS for fermentation, with an additional health benefit from buckwheat.

Amylolytic Activity of Brown Rice and Black Rice during Germination (현미와 흑미의 발아과정 중 amylolytic activity)

  • Lee, Hyang-Mi;Im, Ji-Soon;Park, Jong-Dae;Kum, Jun-Seok;Lee, Hyun-Yu;Lee, Young-Tack
    • Korean Journal of Food Science and Technology
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    • v.45 no.3
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    • pp.333-338
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    • 2013
  • Enzymatic activity in germinated cereal grains is important for the saccharification of starch materials. This study was conducted to investigate the amylolytic activities of germinating brown rice and black rice that have different amylose contents. Brown rice and black rice were steeped at room temperature for 24 h and germinated at 20 and $30^{\circ}C$ for 1, 2, and 3 days. ${\alpha}$- and ${\beta}$-Amylase activities in normal brown rice increased very slightly during the 3-day germination period, but the enzymatic activities were slightly higher in low-amylose (waxy type) brown rice. Diastatic power (DP), a measure of starch-saccharifying enzyme, was higher in the germinating brown rice with low amylose than in those with normal amylose content. ${\alpha}$- and ${\beta}$-Amylase activities in black rice increased gradually during germination, and DP of low-amylose black rice appeared to be higher than that of normal brown rice. Amylase activities in brown rice and black rice germinated at $30^{\circ}C$ were higher than those germinated at $20^{\circ}C$. Compared to brown rice, the overall amylolytic activity of germinated black rice was observed to be higher than that of brown rice.

Optimization of Makgeolli Manufacture Using Several Sweet Potatoes (다양한 고구마를 이용하여 제조한 막걸리의 최적화)

  • Cheon, Ji-Eun;Baik, Moo-Yeol;Choi, Sung-Won;Kim, Chang-Nam;Kim, Byung-Yong
    • The Korean Journal of Food And Nutrition
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    • v.26 no.1
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    • pp.29-34
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    • 2013
  • The objective of this study was to manufacture three kinds of domestic sweet potato Makgeolli using a mixture design and an optimization technique. The effects of four different manufacture methods, such as simultaneous saccharification and fermentation (SSF) with or without malt and separate hydrolysis and fermentation (SHF) with or without malt were determined. The SSF methods of Makgeolli produced higher alcohol content than that of SHF methods. The sensory score was not influenced by different making methods. Fourteen experimental points were selected, and rice (10~50%), sweet potato (10~50%) and water (40~60%) were chosen as independent variables. The measured responses were sensory preference, total polyphenol content, and DPPH radical scavenging activities. The ratio of the optimum sweet potato Makgeolli mixture formulation was developed as 15.11 (rice): 44.89 (sweet potato): 40 (water) using the optimization technique. The desirability of the optimum mixture formulation was 0.839. Yellow sweet potato Makgeolli using the optimum mixture formulation produced higher soluble sugar content compared to others. Regular sweet potato Makgeolli produced higher pH. The purple sweet potato Makgeolli's total polyphenol content and DPPH radical scavenging activity were measured to be the highest at $771.91{\pm}1.42mg\;GAE/{\ell}$, $131.55{\pm}4.03%$.

Enzyme Activities and Cellulose Degradation of Domestic Softwoods in Shaking Culture of Fomitopsis palustris (국내산 침엽수 목분의 진탕배양에서 나타난 Fomitopsis palustris의 효소 활성 및 셀룰로오스 분해)

  • Choi, Doo-Yeol;Lee, Young-Min;Kim, Young-kyoon;Yoon, Jeong-Jun;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.6
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    • pp.91-99
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    • 2007
  • Activities of the extracellular enzyme from Fomitopsis palustris, a brown-rot fungi, and by which crystallinity changes of cellulose in the various softwoods, such as Larix leptolepsis, Finns rigida, Finns koraiensis and Finns densiflora by liquid culture, were investigated. Activity of Cellobiohydrolase (CBH) from F. palustris was detected in the every test softwoods culture, showing activities of the Endoglucanase (EG), $\beta$-glucosidase (BGL) and $\beta$-1,4-xylosidase (BXL). It was shown high enzyme activities in the sapwood culture than heartwood of the same wood species, However, the enzyme activities in most of test wood cultures increased with longer incubation time, indicating a possibility of intermix sapwood and heartwood for degradation process by enzyme. Also it was shown that protein patterns of the extracellular enzyme from F. palustris in wood particle substrate of the several domestic softwoods were similar with each other wood species, which suggested the possibility of mixing all softwoods in saccharification by enzyme from F. palustris. Crystallinity reduction value of cellulose by F. palustris was 4.2~20.4% in 4 weeks cultivation, 12.9~28.9% in 8 weeks.

Effect of alkali pretreatment on bioconversion of waste money bill to glucose for bio-ethanol production

  • Sheikh, M. Mominul Islam;Kim, Chul-Hwan;Park, Hyun-Jin;Kim, Sung-Ho;Kim, Gyeong-Chul;Lee, Ji-Yong;Kim, Jae-Won
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2011.10a
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    • pp.167-177
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    • 2011
  • Renewable energy resources and technologies have the potential to provide long-lasting solutions of the global energy-requirements faced by the economic and environmental sectors of a nation. Therefore, waste money bills were used as renewable energy source for the production of bio-ethanol. In this study, different concentrated NaOH 0.5%. 1.0%, 2.0%, 3.0% and 0.0% (as a control) were used for 10, 20 and 30 mins at $121^{\circ}C$/15 psi in an autoclave. Saccharification and fermentation (aerobic and anaerobic) were carried out through commercial enzyme Celluclast 1.5 L, Novozymes 188 and Saccharomyces cerevisiae KCCM 11304 respectively. The results of pretreatment showed that the NaOH pre-treated substrate enhanced enzyme action and released more amount of glucose. The amount of glucose was found with the increasing concentration of NaOH and time $44996.95{\pm}6.30$, $46763.10{\pm}3.56$, $53421.32{\pm}4.72$, $63431.25{\pm}6.95$ and $56850.98{\pm}6.75\;ng/{\mu}l$ for 30 min respectively. As for bioethanol, the conversion rate of NaOH resulted $1010.08{\pm}4.71$, $1050.25{\pm}4.37$, $1109.49{\pm}4.39$, $1139.25{\pm}3.26$ and $1020.77{\pm}3.89$ ppm for aerobic; $16730.54{\pm}6.67$, $17076.45{\pm}6.25$, $17516.17{\pm}4.49$, $19782.68{\pm}6.19$ and $17973.39{\pm}7.50$ ppm for anaerobic and $18935.02{\pm}4.59$, $19895.45{\pm}5.39$, $21912.95{\pm}4.83$, $24895.21{\pm}6.72$ and $18961.21{\pm}4.90$ ppm for anaerobic condition with benzoic acid for respective condition. Thus, the results of the present work clearly revealed that with the increasing of alkali concentration might be more effective for bio-ethanol production from waste money bill, which is economic and environmental friendly.

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Novel substrate specificity of a thermostable β-glucosidase from the hyperthermophilic archaeon, Thermococcus pacificus P-4 (초고온 고세균 Thermococcus pacificus P-4로부터 내열성 β-glucosidase의 새로운 기질 특이성)

  • Kim, Yun Jae;Lee, Jae Eun;Lee, Hyun Sook;Kwon, Kae Kyoung;Kang, Sung Gyun;Lee, Jung-Hyun
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.68-74
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    • 2015
  • Based on the genomic analysis of Thermococcus pacificus P-4, we identified a putative GH1 ${\beta}$-glucosidase-encoding gene (Tpa-glu). The gene revealed a 1,464 bp encoding 487 amino acid residues, and the deduced amino acid residues exhibited 77% identity with Pyrococcus furiosus ${\beta}$-glucosidase (accession no. NP_577802). The gene was cloned and expressed in Escherichia coli system. The recombinant protein was purified by metal affinity chromatography and characterized. Tpa-Glu showed optimum activity at pH 7.5 and $75^{\circ}C$, and thermostability with a half life of 6 h at $90^{\circ}C$. Tpa-Glu exhibited hydrolyzing activity against various pNP-glycopyranosides, with kcat/Km values in the order of pNP-${\beta}$-glucopyranoside, pNP-${\beta}$-galactopyranoside, pNP-${\beta}$-mannopyranoside, and pNP-${\beta}$-xylopyranoside. In addition, the enzyme exhibited exo-hydrolyzing activity toward ${\beta}$-1,3-linked polysaccharide (laminarin) and ${\beta}$-1,3- and ${\beta}$-1,4-linked oligosaccharides. This is the first description of an enzyme from hyperthermophilic archaea that displays exo-hydrolyzing activity toward ${\beta}$-1,3-linked polysaccharides and could be applied in combination with ${\beta}$-1,3-endoglucanase for saccharification of laminarin.

Bioethanol Production from Wasted Corn Stalk from Gangwon Province : from Enzymatic Hydrolysis to Fermentation (강원지역 폐옥수수대로부터 바이오에탄올 생산 : 효소 당화부터 발효까지)

  • Choi, Jae Min;Choi, Suk Soon;Yeom, Sung Ho
    • Applied Chemistry for Engineering
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    • v.23 no.3
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    • pp.326-332
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    • 2012
  • Among the samples prepared by various pre-treatment methods, the one pretreated by dilute sulfuric acid showed the highest glucose yield in the enzymatic hydrolysis. Statistical analysis of enzymatic hydrolysis revealed that the glucose yield was in proportion to the enzyme dosage, the ratio of the pre-treated sample to the buffer solution, and the reaction time and that the effect of enzyme dosage was predominant in the experiment range. In addition, the glucose yield was estimated to be 76.1% at an optimal enzymatic hydrolysis condition. In a separate hydrolysis and fermentation (SHF), Saccharomyces cerevisiae converted over 80% of glucose from the enzymatic hydrolysis of pre-treated wasted corn stalk by dilute sulfuric acid to bioethanol with 37% of ethanol yield and 0.42 $g/L{\cdot}hr$ of productivity. In the simultaneous saccharification and fermentation (SSF), 59.5% of conversion from glucan to ethanol and 0.20 $g/L{\cdot}hr$ of productivity were achieved. In both SHF and SSF, approximately 88 g of bioethanol could be obtained from 1 kg of wasted corn stalk. The possible amount of bioethanol in Gangwon province were estimated to be 1.9 kiloton with the assumption of the 50% of collection ratio.