scholarly journals Thermostable Xylanase Production by Geobacillus sp. Strain DUSELR13, and Its Application in Ethanol Production with Lignocellulosic Biomass

2018 ◽  
Vol 6 (3) ◽  
pp. 93 ◽  
Author(s):  
Mohit Bibra ◽  
Venkat Kunreddy ◽  
Rajesh Sani
Keyword(s):  
Ethanol Production ◽  
Lignocellulosic Biomass ◽  
Maximum Yield ◽  
Xylanase Activity ◽  
Deep Biosphere ◽  
Xylanase Production ◽  
Beechwood Xylan ◽  
Higher Temperature ◽  
Characterization Studies ◽  
Ph Range

The aim of the current study was to optimize the production of xylanase, and its application for ethanol production using the lignocellulosic biomass. A highly thermostable crude xylanase was obtained from the Geobacillus sp. strain DUSELR13 isolated from the deep biosphere of Homestake gold mine, Lead, SD. Geobacillus sp. strain DUSELR13 produced 6 U/mL of the xylanase with the beechwood xylan. The xylanase production was improved following the optimization studies, with one factor at a time approach, from 6 U/mL to 19.8 U/mL with xylan. The statistical optimization with response surface methodology further increased the production to 31 U/mL. The characterization studies revealed that the crude xylanase complex had an optimum pH of 7.0, with a broad pH range of 5.0–9.0, and an optimum temperature of 75 °C. The ~45 kDa xylanase protein was highly thermostable with t1/2 of 48, 38, and 13 days at 50, 60, and 70 °C, respectively. The xylanase activity increased with the addition of Cu+2, Zn+2, K+, and Fe+2 at 1 mM concentration, and Ca+2, Zn+2, Mg+2, and Na+ at 10 mM concentration. The comparative analysis of the crude xylanase against its commercial counterpart Novozymes Cellic HTec and Dupont, Accellerase XY, showed that it performed better at higher temperature, hydrolyzing 65.4% of the beechwood at 75 °C. The DUSEL R13 showed the mettle to hydrolyze, and utilize the pretreated, and untreated lignocellulosic biomass: prairie cord grass (PCG), and corn stover (CS) as the substrate, and gave a maximum yield of 20.5 U/mL with the untreated PCG. When grown in co-culture with Geobacillus thermoglucosidasius, it produced 3.53 and 3.72 g/L ethanol, respectively with PCG, and CS. With these characteristics the xylanase under study could be an industrial success for the high temperature bioprocesses.

scholarly journals Development of a flat sheet woven fabric membrane fermenter for xylanase production by Thermomyces lanuginosus

2015 ◽  
Author(s):  
◽  
Venessa Thorulsley
Keyword(s):  
Enzyme Activity ◽  
Nutrient Medium ◽  
Shake Flask ◽  
Xylanase Activity ◽  
Membrane System ◽  
Continuous Operation ◽  
Volumetric Productivity ◽  
Xylanase Production ◽  
Flat Sheet ◽  
Beechwood Xylan

Fermentation processes are vital for the production of numerous bioproducts. Fermentation being the mass culture of micro – organisms for the production of some desired product, is an extensive field, with immense prospects for study and improvement. Enzyme production is of significance as these proteins are biological catalysts, finding niches in numerous industries, xylanase for example is utilized in the pulp and paper, animal feed, biofuel and food production processes. During enzyme production, a critical step is biomass separation, whereby the valuable product, the enzyme, is removed from the broth or micro – biological culture before it is denatured. This is typically achieved via centrifugation. The aim of this study was to develop and evaluate a submerged membrane fermenter system with the specific outcome of increasing the rate of production of xylanase, from the thermophilic fungal species Thermomyces lanuginous DSM 5826. Preliminary shake flask experiments were performed to determine the optimal production conditions, followed by partial characterization of the enzyme. A bioreactor was then fabricated to include a flat sheet membrane module, with outlets for permeate and broth withdrawal and inlets for feed and sterile air input. Experiments were conducted to determine the optimal dilution rate for maximum volumetric productivity. Results from the shake flask experiments indicated that the best conditions for xylanase production, yielding xylanase activity of 5118.60 ± 42.76 U.mL-1 was using nutrient medium containing beechwood xylan (1.5 % w/v), yeast extract (1.5 % w/v), potassium dihydrogen phosphate (0.5 % w/v), adjusted to a pH of 6.5 and inoculated with 1.0 mL of spore solution, rotating in a shaking incubator set to 150 rpm at 50 °C. Apart from analysis of the effect of the carbon source on xylanase activity, coarse corn cobs were used in the shake flask experiments as a cost saving initiative. The pH optima was determined to be 6.5 while the temperature optima of the enzyme was 70 °C. SDS PAGE analysis revealed that the molecular weight of the enzyme was between 25 and 35 kDa and qualitative analysis via a zymogram revealed clear zones of hydrolysis on a xylan infused agarose gel. During short run membrane fermenter experiments the percentage increase in enzyme activity between the batch operation (610.58 ± 34.54 U.mL-1) and semi – continuous operation (981.73 ± 55.54 U.mL-1) with beechwood xylan nutrient replenishment was 60.78 %. The maximum volumetric productivity achieved with beechwood supplementation after 192 hours in semi – continuous operation (5.32 ± 0.30 U.mL-1.hr-1) was 2.1 times greater than that of batch operation (2.54 ± 0.14 U.mL-1.hr-1) which equates to an increase of 110.28 % in productivity measured at its peak. The increase in total activity between batch (610 576.92 U) and beechwood xylan medium supplemented semi – continuous mode (1 184 937.50 U) resulted in a 94.07 % increase. During long run experimental periods, the increase in production of xylanase between the batch (873.26 ± 61.78 U.mL-1) and the xylan medium membrane system (1522.41 ± 107.65 U.mL-1) was determined to be 74.34 % while an overall average increase in productivity between the batch and xylan fed membrane system was 43.25%. The total enzyme activity with in membrane mode with beechwood xylan nutrient medium feed was 160 % greater than the batch process offering a 2.6 – fold increase. Experiments where de – ionized water was alternated with beechwood xylan nutrient medium had no significant impact on the productivity or enzyme activity. The optimal dilution rate for maximum volumetric productivity as determined to be 0.0033 hr-1. The results are indicative of the potential viability of such a design, yielding the desired outcome of a membrane integrated system to significantly increase the production of enzymes during fermentation.

Optimization of Xylanase Production by Streptomyces costaricanus 45I-3 Using Various Substrates through Submerged Fermentation

2020 ◽  
Vol 14 (1) ◽  
pp. 5
Author(s):  
SIPRIYADI SIPRIYADI ◽  
ARIS TRI WAHYUDI ◽  
MAGGY THENAWIDJAYA SUHARTONO ◽  
ANJA MERYANDINI
Keyword(s):  
Nitrogen Source ◽  
Yeast Extract ◽  
Xylanase Activity ◽  
Hydrolytic Enzymes ◽  
Tween 80 ◽  
Medium Composition ◽  
Production Costs ◽  
Xylanase Production ◽  
Beechwood Xylan

Xylanase is an important hydrolytic enzymes with many application in several industries, but to obtain enzyme derived products is not easy. Thus, the optimization of efficient xylanases production is a great interest for biotechnological application. This study aims to determine the type of substrate, medium composition, and optimum conditions of xylanase production by S. costaricanus 45I-3. Determination of substrate type was done by growing the tested bacteria on birchwood xylan, beechwood xylan, oat spelled xylan, corn cobs xylan, and tobacco xylan substrate, meanwhile the determination of medium composition and enzyme production were done by measuring xylanase activity at various substrate concentration and replacing the carbon, nitrogen, phosphate and surfactants source. The results showed that the highest enzymatic index (EI) produced from corn cob xylan substrate at 3.60 meanwhile the second highest was beechwood xylan substrate at 2.87 EI, however this substrate is purer, thus this substrate was selected and used as xylan sources for further optimization measurement. The best xylanase activity (2.29 U/mL) obtained on eighth day after inoculation on rotary incubator at 120 rpm in 28 ºC. Arabinose as the source of carbon generate the highest activity at 3.161 U/mL meanwhile the most preferred source of phosphate is Na2HPO4 (2.37 U/mL). Both source of nitrogen i.e. nitrogen ammonium sulphate (NH4)2SO4 and yeast extract were able to produce xylanase at 2.57 and 2.36 U/mL. The addition of surfactant in production medium showed addition of SDS surfactant (0.146 U/mL) and Tween 80 (0.438 U/mL) showed a negative response by decreasing the activity. The conclusion showed that the xylanase activity was increased after optimization at various C, N, and P sources, and the use of nitrogen source (NH4)2SO4), become a more economical alternative to replacing a nitrogen source yeast extract so it can lower the production costs of xylanase enzyme.

scholarly journals Cost-Effective Production and Optimization of Alkaline Xylanase by Indigenous Bacillus mojavensis AG137 Fermented on Agricultural Waste

2011 ◽  
Vol 2011 ◽  
pp. 1-9 ◽  
Author(s):  
Abbas Akhavan Sepahy ◽  
Shokoofeh Ghazi ◽  
Maryam Akhavan Sepahy
Keyword(s):  
Agricultural Waste ◽  
Xylanase Activity ◽  
Nitrogen Sources ◽  
Cost Effective ◽  
Inoculum Size ◽  
Alkaline Condition ◽  
Xylanase Production ◽  
Bacillus Mojavensis ◽  
Oat Bran ◽  
Higher Temperature

A xylanase producer Bacillus mojavensis strain, called AG137, isolated from cotton farm (Kashan-Iran). The optimal xylanase activity reached at 55∘C & pH 9.0. Enzyme yield was studied using a medium with different agricultural wastes as inducers. Xylanase production of about 249.308 IU/mL was achieved at pH 8 and 37∘C, within 48 h submerged fermentation in enzyme production medium supplemented with 2% (w/v) oat bran as an optimum carbon source. A mixture of 1% (w/v) yeast extract and 1% (w/v) tryptone as optimum nitrogen sources, agitation speed 200 rpm, and inoculum size 2% (v/v) were the optimums for maximum production. Accordingly, xylanase yield from 194.68 IU/mL under non-optimized fermentation condition enhanced to 302.466 IU/mL in optimized condition. Screened xylanase is thermostable, presenting 70% stability at 60∘C during 30 min. Further enzyme incubation in higher temperature caused a decrease in the residual enzyme activity, yet it retained 68%–50% of its activity after 1 hour from 45∘C to 55∘C. Besides, it is stable in pH 9 and 10, maintaining over 70% of its activity for 2 h. The enzyme also could preserve 71% and 63% of its initial activity after 3 hours of pre-incubation in the same alkaline condition. Produced xylanase therefore was introduced as an alkaline-active and stable one, displaying suitable thermostability feature, confirmed by HPLC analysis. Hence, all xylanase properties highlight its promising uses in industrial scale.

Optimization of Xylanase Production by Trichoderma reesei JL-1 Using Response Surface Methodology and the Enzymatic Properties

2011 ◽  
Vol 138-139 ◽  
pp. 1183-1189
Author(s):  
Ming Qi Liu ◽  
Xian Jun Dai ◽  
Guang Fu Liu ◽  
Rong Fa Guan
Keyword(s):  
Response Surface Methodology ◽  
Moisture Content ◽  
Response Surface ◽  
Ammonium Sulfate ◽  
Trichoderma Reesei ◽  
Xylanase Activity ◽  
Solid Substrate ◽  
Basic Medium ◽  
Xylanase Production ◽  
Ph Range

The production of xylanase (TrxA) byTrichoderma reeseiJL-1 in solid-state fermentation (SSF) was optimized by response surface methodology (RSM). Results revealed that factors of concentration of added ammonium sulfate, and moisture content had significant effect on the TrxA production (P<0.05). The maximum xylanase activity (485.0 U/g U/g dry fermentation product) was obtained at 2.9% the ammonium sulfate by employing wheat bran as the solid substrate, 61.6% moisture content and 59.5-h fermentation, which was close to the predicted one, and was 1.8 times as high as that of the basic medium. The optimum temperature and pH for TrXA activity were 45°C and pH 5.0, respectively. Over 90% of xylanase activity was retained after treatment of the enzyme by preincubation over a pH range of 3.0-5.0 for 1 h at 25°C. TrxA exhibitedKmandVmaxvalues of 1.458mg/mL and 25.316 μmol/min/ml, respectively. In the presences of metal ions such as Mn2+the activity of the enzyme increased. Whereas strong inhibition of the enzyme activity was observed in the presences of Fe3+and Ca2+.

scholarly journals Method for the Production and Purification of Plant Immuno-Active Xylanase from Trichoderma

2021 ◽  
Vol 22 (8) ◽  
pp. 4214
Author(s):  
Gautam Anand ◽  
Meirav Leibman-Markus ◽  
Dorin Elkabetz ◽  
Maya Bar
Keyword(s):  
Immune System ◽  
Plant Defense ◽  
Plant Immunity ◽  
Xylanase Activity ◽  
Hydrolytic Enzymes ◽  
Adaptive Immune System ◽  
Defense Responses ◽  
Plant Defense Responses ◽  
Xylanase Production ◽  
Ideal System

Plants lack a circulating adaptive immune system to protect themselves against pathogens. Therefore, they have evolved an innate immune system based upon complicated and efficient defense mechanisms, either constitutive or inducible. Plant defense responses are triggered by elicitors such as microbe-associated molecular patterns (MAMPs). These components are recognized by pattern recognition receptors (PRRs) which include plant cell surface receptors. Upon recognition, PRRs trigger pattern-triggered immunity (PTI). Ethylene Inducing Xylanase (EIX) is a fungal MAMP protein from the plant-growth-promoting fungi (PGPF)–Trichoderma. It elicits plant defense responses in tobacco (Nicotiana tabacum) and tomato (Solanum lycopersicum), making it an excellent tool in the studies of plant immunity. Xylanases such as EIX are hydrolytic enzymes that act on xylan in hemicellulose. There are two types of xylanases: the endo-1, 4-β-xylanases that hydrolyze within the xylan structure, and the β-d-xylosidases that hydrolyze the ends of the xylan chain. Xylanases are mainly synthesized by fungi and bacteria. Filamentous fungi produce xylanases in high amounts and secrete them in liquid cultures, making them an ideal system for xylanase purification. Here, we describe a method for cost- and yield-effective xylanase production from Trichoderma using wheat bran as a growth substrate. Xylanase produced by this method possessed xylanase activity and immunogenic activity, effectively inducing a hypersensitive response, ethylene biosynthesis, and ROS burst.

Ethanol Production from Lignocellulosic Biomass Using Xylotrophic Basidiomycetes

2015 ◽  
Vol 51 (5) ◽  
pp. 516-525 ◽  
Author(s):  
N. R. Al’myasheva ◽  
A. A. Novikov ◽  
E. Yu. Kozhevnikova ◽  
A. V. Golyshkin ◽  
A. V. Barkov ◽  
...  
Keyword(s):  
Ethanol Production ◽  
Lignocellulosic Biomass ◽  
Xylotrophic Basidiomycetes

scholarly journals Synergistic Effect of Different Plant Cell Wall–Degrading Enzymes Is Important for Virulence of Fusarium graminearum

2017 ◽  
Vol 30 (11) ◽  
pp. 886-895 ◽  
Author(s):  
Maria Chiara Paccanaro ◽  
Luca Sella ◽  
Carla Castiglioni ◽  
Francesca Giacomello ◽  
Ana Lilia Martínez-Rocha ◽  
...  
Keyword(s):  
Cell Wall ◽  
Fusarium Graminearum ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Xylanase Activity ◽  
Cellulase Activity ◽  
Wild Type ◽  
Cell Wall Degrading Enzymes ◽  
Xylanase Production ◽  
Significant Difference

Endo-polygalacturonases (PGs) and xylanases have been shown to play an important role during pathogenesis of some fungal pathogens of dicot plants, while their role in monocot pathogens is less defined. Pg1 and xyr1 genes of the wheat pathogen Fusarium graminearum encode the main PG and the major regulator of xylanase production, respectively. Single- and double-disrupted mutants for these genes were obtained to assess their contribution to fungal infection. Compared with wild-type strain, the ∆pg mutant showed a nearly abolished PG activity, slight reduced virulence on soybean seedlings, but no significant difference in disease symptoms on wheat spikes; the ∆xyr mutant was strongly reduced in xylanase activity and moderately reduced in cellulase activity but was as virulent as wild type on both soybean and wheat plants. Consequently, the ΔpgΔxyr double mutant was impaired in xylanase, PG, and cellulase activities but, differently from single mutants, was significantly reduced in virulence on both plants. These findings demonstrate that the concurrent presence of PG, xylanase, and cellulase activities is necessary for full virulence. The observation that the uronides released from wheat cell wall after a F. graminearum PG treatment were largely increased by the fungal xylanases suggests that these enzymes act synergistically in deconstructing the plant cell wall.

scholarly journals Ethanol production using vegetable peels medium and the effective role of cellulolytic bacterial (Bacillus subtilis) pre-treatment

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 271
Author(s):  
Salman Khan Promon ◽  
Wasif Kamal ◽  
Shafkat Shamim Rahman ◽  
M. Mahboob Hossain ◽  
Naiyyum Choudhury
Keyword(s):  
Bacillus Subtilis ◽  
Ethanol Production ◽  
Energy Demand ◽  
Biochemical Characterization ◽  
Raw Materials ◽  
Maximum Yield ◽  
Clean Energy ◽  
Raw Material ◽  
Cellulolytic Bacteria ◽  
Alcohol Production

Background: The requirement of an alternative clean energy source is increasing with the elevating energy demand of modern age. Bioethanol is considered as an excellent candidate to satiate this demand.Methods:Yeast isolates were used for the production of bioethanol using cellulosic vegetable wastes as substrate. Efficient bioconversion of lignocellulosic biomass into ethanol was achieved by the action of cellulolytic bacteria (Bacillus subtilis).  After proper isolation, identification and characterization of stress tolerances (thermo-, ethanol-, pH-, osmo- & sugar tolerance), optimization of physiochemical parameters for ethanol production by the yeast isolates was assessed. Very inexpensive and easily available raw materials (vegetable peels) were used as fermentation media. Fermentation was optimized with respect to temperature, reducing sugar concentration and pH.Results:It was observed that temperatures of 30°C and pH 6.0 were optimum for fermentation with a maximum yield of ethanol. The results indicated an overall increase in yields upon the pretreatment ofBacillus subtilis; maximum ethanol percentages for isolate SC1 obtained after 48-hour incubation under pretreated substrate was 14.17% in contrast to untreated media which yielded 6.21% after the same period. Isolate with the highest ethanol production capability was identified as members of the ethanol-producingSaccharomycesspecies after stress tolerance studies and biochemical characterization using Analytical Profile Index (API) ® 20C AUX and nitrate broth test. Introduction ofBacillus subtilisincreased the alcohol production rate from the fermentation of cellulosic materials.Conclusions:The study suggested that the kitchen waste can serve as an excellent raw material in ethanol fermentation.

scholarly journals Bio-ethanol (2nd Generation Ethanol): A Solution to Ever Polluting Gasoline to Climate in India

2020 ◽  
pp. 1-15
Author(s):  
Shruti Mohapatra ◽  
Raj Kishore Mishra ◽  
Khitish K. Sarangi
Keyword(s):  
Ethanol Production ◽  
Lignocellulosic Biomass ◽  
Fossil Fuels ◽  
Economic Benefits ◽  
Policy Implications ◽  
Current Status ◽  
Environmentally Sustainable ◽  
2Nd Generation ◽  
Production Technologies ◽  
Rapid Pace

Environmentally sustainable energy sources are called for due to contemporaneous development in industries along with the rapid pace of urbanization. Ethanol produced from biomass can be deliberated as a clean and safest liquid fuel and an alternative to fossil fuels as they have provided unique environmental, strategic economic benefits. For the past decade, it has been noticed that there is an increasing trend found in bio ethanol production which has created a stimulus to go for advancement in bio ethanol production technologies. Several feed stocks have been used for the bio ethanol production but the second generation bio ethanol has concentrated on the lignocellulosic biomass. Plenteous lignocellulosic biomass in the world can be tapped for ethanol production, but it will require significant advances in the ethanol production process from lignocellulosic because of some technical and economic hurdles found in commercial scale. This review will encompass the current status of bio ethanol production in terms of their economic and environmental viability along with some research gaps as well as policy implications for the same.

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