• Title/Summary/Keyword: Cellulosic biomass

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Comprehensive Characterization of Mutant Pichia stipitis Co-Fermenting Cellobiose and Xylose through Genomic and Transcriptomic Analyses

  • Dae-Hwan Kim;Hyo-Jin Choi;Yu Rim Lee;Soo-Jung Kim;Sangmin Lee;Won-Heong Lee
    • Journal of Microbiology and Biotechnology
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    • v.32 no.11
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    • pp.1485-1495
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    • 2022
  • The development of a yeast strain capable of fermenting mixed sugars efficiently is crucial for producing biofuels and value-added materials from cellulosic biomass. Previously, a mutant Pichia stipitis YN14 strain capable of co-fermenting xylose and cellobiose was developed through evolutionary engineering of the wild-type P. stipitis CBS6054 strain, which was incapable of co-fermenting xylose and cellobiose. In this study, through genomic and transcriptomic analyses, we sought to investigate the reasons for the improved sugar metabolic performance of the mutant YN14 strain in comparison with the parental CBS6054 strain. Unfortunately, comparative whole-genome sequencing (WGS) showed no mutation in any of the genes involved in the cellobiose metabolism between the two strains. However, comparative RNA sequencing (RNA-seq) revealed that the YN14 strain had 101.2 times and 5.9 times higher expression levels of HXT2.3 and BGL2 genes involved in cellobiose metabolism, and 6.9 times and 75.9 times lower expression levels of COX17 and SOD2.2 genes involved in respiration, respectively, compared with the CBS6054 strain. This may explain how the YN14 strain enhanced cellobiose metabolic performance and shifted the direction of cellobiose metabolic flux from respiration to fermentation in the presence of cellobiose compared with the CBS6054 strain.

Characterization of Cellulases from Schizophyllum commune for Hydrolysis of Cellulosic Biomass (Schizophyllum commune에 의한 Cellulase 생산 및 섬유소계 바이오매스의 당화를 위한 효소적 특성)

  • Kim, Hyun-Jung;Kim, Yoon-Hee;Cho, Moon-Jung;Shin, Keum;Lee, Dong-Heub;Kim, Tae-Jong;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.6
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    • pp.547-560
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    • 2010
  • The optimum culture condition of Schizophyllum commune for the cellulase production and its enzymatic characteristics for saccharification of cellulosic biomass were analyzed. S. commune secrets ${\beta}$-1,4-xylosidase (BXL) and cellulases, including endo-${\beta}$-1,4-glucanase (EG), cellobiohydrolase (CBH), and ${\beta}$-glucosidase (BGL). The optimum reaction temperature for all cellulases was $50^{\circ}C$ and the thermostable range was $30{\sim}40^{\circ}C$C. The optimum reaction pH for all cellulases was 5.5 in a range of temperature from $0^{\circ}C$ to $55^{\circ}C$. The best nutritions for the cellulase production of S. commune among tested nutrients were 2% cellulose for the carbon source and corn steep liquor or peptone/yeast extract for the nitrogen source without vitamins. The environmental culture condition for the cellulase production was 5.5~6.0 for pH at $25{\sim}30^{\circ}C$. The enzyme activities of EG, BGL, CBH, and BXL were 3670.5, 631.9, 398.5, and 15.2 U/$m{\ell}$, respectively, after concentration forty times from the culture broth of S. commune which was grown at the optimized culture condition. Alternative filter paper unit assay showed 11 FPU/$m{\ell}$ enzyme activity. The saccharification tests using cellulase of S. commune showed the low saccharification rate on tested hardwoods but a high value of 50.5% on cellulose, respectively. The saccharification rate (50.5%) of cellulose by cellulase produced in this work is higher than 45.7% in the commercial enzyme (Celluclast 1.5L, 30 FPU/g, glucan).

Growth of Bioenergy Crop Miscanthus sacchariflorus cv. Geodae 1 on Barren Reclaimed Land Applied with Solidified Sewage Sludge in Landfill Sites (하수슬러지 고화물을 처리한 유휴 간척지토양에서 바이오에너지작물 거대 1호의 생육특성 연구)

  • An, Gi Hong;Jang, Yun-Hui;Um, Kyoung Ran;Yu, Gyeong-Dan;Lee, Ji-Eun;Cha, Young-Lok;Moon, Yun-Ho;Ahn, Jong Woong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.3
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    • pp.374-380
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    • 2015
  • This study firstly provides basic data for selection of cultivatable bioenergy grass in barren reclaimed lands applied with solidified sewage sludge. The experimental plots consisted of a plot containing reclaimed land mixed with solidified sewage sludge (MSS 50), a plot covered by solidified sewage sludge (CSS 100), and an original reclaimed soil plot (ORS). The growth, biomass production of bioenergy grasses and soil chemical properties were investigated in each experimental plot for 5 years. The organic matter (OM) and total nitrogen (T-N) content in both MSS 50 and CSS 100 were considerably higher than those in ORS. In bioenergy grasses, M. sacchariflorus cv. Geodae 1 showed an excellent growth and adaptability on reclaimed land applied with solidified sewage sludge. The application of solidified sewage sludge may provided soil nutrition in the reclaimed land due to the fact that bioenergy crops grew better in soils applied with solidified sewage sludge than in untreated soils, and treated soils had higher OM and T-N content than untreated soils. This study suggests that M. sacchariflorus cv. Geodae 1 is the most suitable biomass feedstock crop for biomass production and that solidified sewage sludge may be used as a soil material for cultivation of bioenergy grass on reclaimed lands.

A Study on the Conversion to Feed Stuff from Cellulosic Biomass (섬유질자원(纖維質資源)의 사료(飼料) 전환(轉換))

  • Lee, Ke-Ho;Sung, Chang-Geun;Chung, Kyu-Ok
    • Applied Biological Chemistry
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    • v.27
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    • pp.29-46
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    • 1984
  • To utilize several species of hard wood as raw materials of feed products, fermentation characteristics of cellulosic substrates to single cell protein was investigated, and results were summarized as follows. Among the microorganisms investigated, Tricoderma viride was selected as one of the most cellulolytic. Mixed culture of fungi did not show a synergistic effect on cellulose degradation. When the fungi were cultured at $28^{\circ}C$ for 7 days in a medium containing wheat bran 25 g, cellulose 0.25 g, proteose peptone 0.025 g and tween 800.025 g, cellulotic activities on carboxy methyl cellulose and filter paper reached maximum at 12 hr. The alkali treatment resulted in increased degradation of substrate from 13 to 18% when treated with enzymes for 12h, and reducing sugar formation increased with decreased size of substrates. Glucose was a very good feedback inhibitor of the enzyme from T.viride than that of xylose. When the substrate was rehydrolyzed, hydrolysis rate was 31% to reducing sugars within 12 hr. Quantative anlysis with HPLC showed the ratio of glucose to xylose in sugar syrups as 1.77 to 1. For the purpose of producing cellulosic-single cell protein from the sawdust of mulberry tree, 15 strains of xylose-assimilating yeast were isolated from 42 samples of rotten woods and compost soils and examined for their ability to utilize xylose. Then three strains were selected by their strong xylose-assimilating activities. The cultivative condition, the growth characteristics, and protein and nucleic acid productivities of three strains were investigated. The results obtained were, 1. Wood hydrolysate of mulberry tree was assimilated by 5 strains of CHS-2, CHS-3, ST-40, CHS-12 and CHS-13. 2. The optimum initial pH and temperature for the growth of strain CHS-13 were 4.4 and $30^{\circ}C$. 3. The specific growth rate of strain CHS-13 was $0.23h^{-1}$ and generation time was 3.01 hrs at the optimum condition. 4. CHS-13 strain assimilated 81 % of sugar in wood hydrolysate. 5. CHS-13 strain was identified as Candida guilliermondii var. guilliermondii 6. When the CHS-13 strain was cultured in the wood hydrolysate containing yeast extract, L-protein content was increased with yeast extract concentration. 7. The L-protein and nucleic acid yields from wood hydrolysate were 0.73 mg/ml and $4.92{\times}10^{-2}\;mg/ml$ respectively. 8. An optimal nucleic acid content of CHS-13 strain was observed in the medium containing 0.2% of yeast extract.

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Characteristics of Membrane Filtration as a Post Treatment to Anaerobic Digestion (혐기성 소화의 후처리로서 분리막의 여과특성 연구)

  • Choo, Kwang-Ho;Lee, Chung-Hak;Pek, Un-Hwa;Koh, Ui-Chan;Kim, Sang-Won;Koh, Jong-Ho
    • Applied Chemistry for Engineering
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    • v.3 no.4
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    • pp.730-738
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    • 1992
  • Filtration characteristics according to membrane materials were studied In the ultrafiltration of anaerobic digestion broth as a post treatment method. A series of resistances for different membranes were quantitatively assessed on the basis of the resistance-in-series model. Flux behavior observed with the digestion broth was irrelevant to initial water permeabilities of each membrane. The fluoro polymer membrane showed the most significant improvement of flux with increase of cross-flow velocity, which suggests that the cake layer formed on this membrane is more weakly attached to the membrane surface than those on the other membranes. Flux reduction during longtime running was attrib-used to the polarization layer resistance ($R_p$) as well as the fouling layer resistance($R_f$). Continuous increase of $R_p$ may reflect the variation in the characteristics of cake layers, which could result from size, shape, and structure changes due to lysis and growth of biomass. Hydrophilic cellulosic membrane had a much lower fouling tendency than hydrophobic polysulfone membrane. The depressurization method induced a small increase in flux of $5-10L/m^2/h$. During washing and cleaning, filtrability of each membrane was rapidly recovered within 15 minutes until a stationary value was reached.

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