Composition of Synthesized Cellulolytic Enzymes Varied with the Usage of Agricultural Substrates and Microorganisms
Siddheshwar D. KshirsagarPankajkumar R. WaghmareGanesh Dattatraya SarataleRijuta Ganesh SarataleMayur B. KuradeByong‐Hun JeonSanjay P. Govindwar
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Keywords:
Phanerochaete
Trichoderma viride
Chrysosporium
Aspergillus awamori
Trichoderma harzianum
Solid-State Fermentation
Phanerochaete
Trichoderma viride
Chrysosporium
Aspergillus awamori
Trichoderma harzianum
Solid-State Fermentation
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Cellulase and xylanase are enzymes produced by microorganisms to breakdown a component of the plant cell walls. Particular fungal strains are interesting enzyme producers due to their higher production level of cellulase and xylanase in comparison with yeast and bacteria. In this study, the potential of the fungal strains Aspergillus awamori BCC13292, A. niger BCC7037, A.flavus BCC18310, Trichoderma harzianum BCC17752, T.reesei BCC7041 and Penicillium chrysogenum TISTR3554, were investigated for their ability to produce enzymes. Cellulase and xylanase production were detected with an agar plate containing carboxymethyl-cellulose (CMC) and xylan as the carbon source, respectively. The results showed that A.niger cultured in CMC, and T.reesei cultured in xylan, revealed the largest clear zones of 1.50 and 3.05 cm., respectively. The cellulase and xylanase activities of these fungal strains cultured in a liquid medium containing CMC and xylan were also assayed. The results showed that A.flavus cultured in xylan exhibited the highest xylanase activity (1.06 U/mg proteins), while the greatest cellulase activity was obtained from T.reesei (6.22 U/mg proteins) cultured in CMC. In addition, each fungal strain was cultured in a liquid medium consisting of corn husks to test their ability in enzyme production. The greatest cellulase and xylanase activities were acquired from A.niger and T.reesei with 0.41 U/mg proteins and 0.67 U/mg proteins, respectively.
Aspergillus niger
Trichoderma harzianum
Carboxymethyl cellulose
Aspergillus awamori
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Solid state fermentation of wheat straw for cellulase production by Phanerochaete chrysosporium has been studied. The most optimum moisture level for solid state fermentation was found to be 40% while the best enzyme yield was obtained at an initial pH of 4.5 and incubation temperature of 30 0 C. Supplementation @ 0.2% (W/W) was found to have enhanced the yield of the enzyme by three folds. With all conditions standardized an enzyme yield of 78 IU pergds could be obtained.
Solid-State Fermentation
Phanerochaete
Chrysosporium
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Two agro-industrial coproducts, soybean cotyledon fiber and distiller's dried grains with solubles (DDGS), were used as substrates to evaluate the effect of coculturing three different fungi, Aspergillus oryzae, Trichoderma reesei, and Phanerochaete chrysosporium, on enzyme production by solid-state fermentation (SSF). When soybean fiber was used as the substrate, a maximum xylanase activity of 757.4 IU/g and a cellulase activity of 3.2 IU/g were achieved with the inoculation and incubation of T. reesei and P. chrysosporium for 36 h, followed by A. oryzae for an additional 108 h. This inoculation scheme also resulted in the highest xylanase activity of 399.2 IU/g compared to other fungi combinations in the SSF of DDGS. A large-scale SSF by this fungus combination produced fermented products that had xylanase and cellulase activities of 35.9-57.0 and 0.4-1.2 IU/g, respectively. These products also had 3.5-15.1% lower fiber and 1.3-4.2% higher protein contents, suggesting a potential feed quality improvement.
Solid-State Fermentation
Coproduct
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목질계 분해효소 활성 증대를 위해 밀짚을 이용한 고체발효에서 주요 발효인자의 최적화를 수행하였다. Trichoderma reesei와 Aspergillus niger를 이용한 혼합배양에서 고체발효에 주요한 영향을 미친다고 알려진 배양온도, pH, 수분함량과 고체기질 크기를 순차적 최적화를 진행하였다. 실험에 적용 된 인자 모두 목질계 분해효소 활성에 유의한 효과를 주었으며, 발효온도 $40^{\circ}C$ , pH 7, 수분함량 75%와 고체기질 크기 0.25~0.5 mm가 목질계 분해효소 생산을 위한 최적 조건임을 알 수 있었다. 최적조건 하에서 밀짚을 이용한 고체발효를 수행하였을 때, 효소활성 기준 cellulase 10.3 IU, endoglucanase 100.3 IU, ${\beta}$ -glucosidase 22.9 IU와 xylanase 2261.7 IU/g dry material을 배양 96시간에 확인할 수 있었다. 본 결과는 기존 효소활성 대비 각각 72.6, 48.7, 55.2와 51.9% 증가한 수치로 혼합배양과 순차적 최적화를 적용하여 효과적인 목질계 분해효소 활성 증대가 가능함을 확인하였다. The present study was carried out to optimize fermentation parameters for the production of cellulolytic enzymes through solid substrate fermentation of Trichoderma reesei and Aspergillus niger grown on wheat straw. A sequential optimization based on one-factor-at-a-time method was applied to optimize fermentation parameters including temperature, pH, moisture content and particle size. The results of optimization indicated that $40^{\circ}C$ , pH 7, moisture content 75% and particle size between 0.25~0.5 mm were found to be the optimum condition at 96 hr fermentation. Under the optimal condition, co-culture of T. reesei and A. niger produced cellulase activities of 10.3 IU, endoglucanase activity of 100.3 IU, ${\beta}$ -glucosidase activity of 22.9 IU and xylanase activity of 2261.7 IU/g dry material were obtained. Cellulolytic enzyme production with optimization showed about 72.6, 48.8, 55.2 and 51.9% increase compared to those obtained from control experiment, respectively.
Aspergillus niger
Solid-State Fermentation
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Abstract Autohydrolyzed and ethanol‐alkali pulped wheat straw was examined as a candidate feedstock for both cellulase and xylanase production and enzymatic hydrolysis. Submerged cultures of Trichoderma reesei F‐522 grown on hydrothermally modified straw provided culture supernatants of the highest enzymatic activities, whereas the maximal efficiency of enzymatic hydrolysis was recorded in straw treated with ethanol‐NaOH mixture. Some culture conditions were optimized to improve the growth and cellulase production by T. reesei on autohydrolyzed wheat straw.
Enzymatic Hydrolysis
Rice straw
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A 28-kDa endoglucanase was isolated from the culture filtrate of Phanerochaete chrysosporium strain K3 and named EG 28. It degrades carboxymethylated cellulose and amorphous cellulose, and to a lesser degree xylan and mannan but not microcrystalline cellulose (Avicel). EG 28 is unusual among cellulases from aerobic fungi, in that it appears to lack a cellulose-binding domain and does not bind to crystalline cellulose. The enzyme is efficient at releasing short fibres from filter paper and mechanical pulp, and acts synergistically with cellobiohydrolases. Its mode of degrading filter paper appears to be different to that of endoglucanase I from Trichoderma reesei. Furthermore, EG 28 releases colour from stained cellulose beads faster than any other enzyme tested. Peptide mapping suggests that it is not a fragment of another known endoglucanases from P. chrysosporium and peptide sequences indicate that it belongs to family 12 of the glycosyl hydrolases. EG 28 is glycosylated. The biological function of the enzyme is discussed, and it is hypothesized that it is homologous to EG III in Trichoderma reesei and the role of the enzyme is to make the cellulose in wood more accessible to other cellulases.
Phanerochaete
Chrysosporium
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In this study, agriculture waste palm empty fruit bunch (EFB) was used as carbon/cellulose source in solid state fermentation for cheaper cellulase production. Fermentation operation parameters, such as: solid to liquid ratio, temperature, and pH, were varied to study the effect of those parameters towards crude cellulase activity. Two different fungi organisms, Trichoderma viride and Trichoderma reesei were used as the producers. Extracellular cellulase enzyme was extracted using simple contact method using citrate buffer. Assessment of the extracted cellulase activity by filter paper assay showed that Trichoderma viride is the superior organism capable of producing higher cellulase amount compared to Trichoderma reesei at the same fermentation condition. The optimum cellulase activity of 0.79 FPU/g dry substrate was obtained when solid to liquid ratio used for the fermentation was 1:1, while the optimum fermentation temperature and pH were found to be 30 °C and 5.5, respectively. The result obtained in this research showed the potential of EFB utilization for enzyme production.
Trichoderma viride
Solid-State Fermentation
Filter paper
Trichoderma
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The enzyme production for lignocellulose saccharification by solid-state fermentation (SSF) of a food manufacturing byproduct was successfully carried out in a 30 L rotary bioreactor. Defatted spent copra (SC) supplemented with wheat bran (WB) was used as a substrate for the SSF of Trichoderma reesei and aerated at various rates. Regression analysis showed that the carbohydrate/protein (C/P) ratio of the substrate and the supplied aeration rate were the important factors for producing the enzyme cocktail, including cellulases (FPase, CMCase, and cellobiase) and xylanase. The substrate containing SC:WB of 3:2 (or the C/P ratio of 5.4) and the aeration of 1.0 L kg–1substrate min–1 were found to enhance the production of the enzymes up to 5.68, 8.66, 29.2, and 34.44 U g–1 of dry substrate for FPase, CMCase, cellobiase, and xylanase activities, respectively. This discovery provided a promising environment for other substrates to produce multi-enzymes for lignocellulosic saccharification. Additionally, mathematical models were generated to predict the saccharifying degree of the produced enzyme for lignocellulose saccharification.
Solid-State Fermentation
Aspergillus awamori
Bioconversion
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