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 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.

scholarly journals Production and Partial Characterization of an Alkaline Xylanase from a Novel FungusCladosporium oxysporum

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Guo-Qiang Guan ◽  
Peng-Xiang Zhao ◽  
Jin Zhao ◽  
Mei-Juan Wang ◽  
Shu-Hao Huo ◽  
...  
Keyword(s):  
Nitrogen Source ◽  
Wheat Bran ◽  
Internal Transcribed Spacer ◽  
Gene Sequence ◽  
Agricultural Waste ◽  
Xylanase Activity ◽  
Maximum Activity ◽  
Alkaline Proteases ◽  
Xylanase Production

A new fungusCladosporium oxysporumGQ-3 producing extracellular xylanase was isolated from decaying agricultural waste and identified based on the morphology and comparison of internal transcribed spacer (ITS) rDNA gene sequence.C. oxysporumproduced maximum xylanase activity of 55.92 U/mL with wheat bran as a substrate and NH4Cl as a nitrogen source. Mg2+improvedC. oxysporumxylanase production.Partially purified xylanase exhibited maximum activity at 50°C and pH 8.0, respectively, and showed the stable activity after 2-h treatment in pH 7.0–8.5 or below 55°C. Mg2+enhanced the xylanase activity by 2% while Cu2+had the highest inhibition ratio of 57.9%. Furthermore,C. oxysporumxylanase was resistant to most of tested neutral and alkaline proteases. Our findings indicated thatCladosporium oxysporumGQ-3 was a novel xylanase producer, which could be used in the textile processes or paper/feed industries.

Optimization of Medium Components for β-Glucosidase Production from Aspergillus Niger by Response Surface Methodology

2013 ◽  
Vol 419 ◽  
pp. 328-333 ◽  
Author(s):  
Chao Zhang ◽  
Rui Huang ◽  
Hui Tian ◽  
Ru Ming Zhao ◽  
Fa Shun Yu ◽  
...  
Keyword(s):  
Response Surface Methodology ◽  
Response Surface ◽  
Yeast Extract ◽  
Influence Factors ◽  
Mutual Effect ◽  
Tween 80 ◽  
Fermentation Medium ◽  
Medium Composition ◽  
Malt Extract ◽  
Medium Components

β-Glucosidase is the key enzyme for the utilization of lignocellulose.But the commercial β-glucosidase can’t be produced. This paper focuses on the study of the β-glucosidase fermentation process.The fermentation medium components for β-glucosidase production from Aspergil lusniger was optimized by response surface methodology (RSM). Firstly, the three of the most important influence factors yeast extract, MnSO4•H2O and MgSO4•7H2O was obtained from Plackett-Burman design screening. Then the path of steepest ascent experiment was adopted to approach the optimal region of the medium composition. Lastly, the optimal concentration and mutual effect of three factors were predicted by RSM. The results showed that the best medium composition was Malt extract 18g/L, Yeast extract 3.22g/L, KH2PO4 3g/L, MnSO4•H2O 0.58mM, Tween-80 0.5mL/L and MgSO4•7H2O 0.23g/L. Under these fermentation conditions, the activity of β-glucosidase was up to 7.33IU/mL with increasing 23.2% than before.

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.

scholarly journals Increasing Chitinase Activity of Serratia marcescens PT-6 through Optimization of Medium Composition

2019 ◽  
Vol 14 (3) ◽  
pp. 113
Author(s):  
Akhmad Awaludin Agustiar ◽  
Imas Faturrohmah ◽  
Bekti Wulan Sari ◽  
Nurul Binti Isnaini ◽  
Indun Dewi Puspita ◽  
...  
Keyword(s):  
Carbon Source ◽  
Serratia Marcescens ◽  
Nitrogen Source ◽  
Yeast Extract ◽  
Value Added ◽  
Chitinase Activity ◽  
Medium Composition ◽  
Colloidal Chitin ◽  
Additional Carbon Source ◽  
Additional Nitrogen

Chitin hydrolysate is one of the value added product derived from shrimp shell waste. Production of chitin hydrolysate using biological process offers an environmental friendly method compared to chemical process. Serratia marcescens PT-6, a gram negative chitinolytic bacterium isolated from shrimp pond sediment, shows good activity in hydrolyzing chitin. This study aimed to improve the chitinase activity of S. marcescens PT-6 culture by optimizing the component of chitin-containing medium (additional nitrogen source, additional carbon source, and colloidal chitin). The optimization of chitinase by S. marcescens PT-6 culture was done using one variable at a time method. The sequence of the research were to optimize 1) the type of additional carbon source (glucose, lactose, sucrose, and starch), 2) the type of additional nitrogen source (yeast extract, peptone, ammonium sulphate, and ammonium chloride), 3) the concentration of colloidal chitin (0.5; 1; 1.5; 2; and 2.5%), and 4) the concentration of the additional carbon and nitrogen source. The culture of S. marcescens PT-6 was incubated in colloidal chitin medium at 30 oC and chitinase activity from culture supernatant was analyzed. The results showed that starch gave the highest chitinase activity compare to other carbon source, meanwhile yeast extract was chosen as the best nitrogen source among others. The combination of 1.5% colloidal chitin with 0.5% starch and 0.1% yeast extract in medium increased the chitinase activity of S. marcescens PT-6 to 0.021 U/ml. These results indicated that an appropriate medium composition could increase the chitinase activity produced by S. marcescens PT-6 culture.

Enhanced Soil Washing of PCB by Sophorolipids from Transformer Oil Contaminated Soil

2014 ◽  
Vol 694 ◽  
pp. 450-454 ◽  
Author(s):  
Xiao Yu Qi ◽  
Xin Song ◽  
You Min Sun
Keyword(s):  
Nitrogen Source ◽  
Removal Efficiency ◽  
Yeast Extract ◽  
Contaminated Soil ◽  
Fermentation Broth ◽  
Soil Washing ◽  
Tween 80 ◽  
Transformer Oil ◽  
Indicator Pcbs ◽  
Synthetic Surfactants

The effects of surfactants including synthetic surfactants and biosurfactants on 7 indicator PCBs removal from transformer oil-contaminated soil by using batch soil washing assessment were investigated. The sophorolipids (SLs) produced by Wickerhamiella domercqiae var. sophorolipid, especially lactonic SLs exhibited much higher PCB removal efficiency than SDS and Tween-80. The Σ7 PCB removal by 1.0 g/L lactonic SLs, 5.0 g/L of total SLs and acidic SLs was 30.2%, 30% and 25.4%, respectively. Removal efficiency of lower chlorinated congeners was significantly increased by adding sophorolipid, but higher chlorinated congeners removal efficiency was not further improved. 0.5 g/L lactonic SLs was efficient for PCB101 and 118 removal while 1.0 g/L lactonic SLs was efficient for PCB28 and 52 removal. The removal percentage was not increased with the increase of lactonic SLs concentration. The fermentation broth (yeast extract as nitrogen source) of W. domercqiae that was used directly on PCB removal was of superior efficiency.

scholarly journals Screening of Culture Conditions for Production of Xylanase from Landfill Soil Bacteria

2019 ◽  
Vol 19 (2) ◽  
pp. 470 ◽  
Author(s):  
Siti Nor Amira Rosli ◽  
Rohaida Che Man ◽  
Nasratun Masngut
Keyword(s):  
Carbon Source ◽  
Moisture Content ◽  
Nitrogen Source ◽  
Incubation Period ◽  
Xylanase Activity ◽  
Nitrogen Sources ◽  
Culture Conditions ◽  
Inoculum Size ◽  
Xylanase Production ◽  
Initial Ph

Culture conditions including initial pH media, incubation period, inoculum size, type of carbon source, type of nitrogen source and its concentration, which affect xylanase production were screened via the one-factor-at-a-time approach. The bacteria used in the production of xylanase was isolated from the landfill site at Sg. Ikan, Kuala Terengganu, Malaysia. Three characterizations of the landfill soil were investigated for their moisture content, ash content, and pH. The culture conditions range used in the experimental work were between 6–30 h for the incubation period, with initial pH between 5–9, inoculum size between 1–20% v/v, carbon, nitrogen sources, and nitrogen source concentration between 1–5% w/v. Xylanase activity was estimated using dinitrosalicylic acid (DNS) based on the release of xylose under standard assay conditions. The landfill soil was observed to have pH between pH 3.4–7.2 with a moisture content between 12.4–33.7% and ash ranged between 3.5–4.3%. Results showed that the highest xylanase activity within studied ranges was recorded at 25.91±0.0641 U/mL with 10% (v/v) inoculum size, 1% (w/v) xylose as sole carbon source, mixture of 1% (w/v) peptone and 0.25% (w/v) ammonium sulphate as nitrogen sources, which was carried out at initial pH of 8.0 for 24 h incubation.

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.

Bioethanol production from alkaline hydrogen peroxide pretreated Populus deltoides wood using hydrolytic enzymes of Bacillus stratosphericus N12(M) and Bacillus altitudinis Kd1(M) under different modes of separate hydrolysis and fermentation by monoculture and co-culture combinations of ethanologens

2016 ◽  
Vol 5 (02) ◽  
pp. 4810
Author(s):  
Nisha Sharma* ◽  
Nivedita Sharma
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Populus Deltoides ◽  
Ethanol Yield ◽  
Hydrolytic Enzyme ◽  
Xylanase Activity ◽  
Hydrolytic Enzymes ◽  
Cellulase Activity ◽  
Integrated Approach ◽  
Xylanase Production ◽  
Bacillus Altitudinis

An integrated approach was studied for in-house cellulase and xylanase production, from novel hyper hydrolytic enzyme producers and enzymatic hydrolysis of pretreated Populus deltoides wood into bioethanol. A xylanase producer Bacillus altitudinis Kd1 (M) and cellulase producerBacillus stratosphericus N12 (M) was isolated from soil. Optimization of process parameters led to an optimal xylanase activity of 96.25 IU at 300C and pH 5.5 and cellulase activity of 5.98 IU at 300C and pH 8.0. The NaOH+H2O2 pretreated biomass was hydrolysed using cellulase and xylanase producing 12.45 mg/g of reducing sugars. Further fermentation of lignocellulosic hydrolysate was performed using different yeasts viz. Saccharomyces cerevisiae I, Saccharomyces cerevisiae II, Pichia stipitis, Candida shehatae and Zymomonas mobilis and maximum 11.10 g/l ethanol yield achieved with co-culture of S. cerevisiae II + P. stipitis with fermentation efficiency of 43.52% under method IV of SHF. The results have significant implications and further applications regarding production of fuel ethanol from agricultural lignocellulosic waste.

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