scholarly journals Impact of cultivation conditions on xylanase production and growth in Paenibacillus mucilaginosus

Author(s):  
D. T. Ha ◽  
A. V. Kanarskiy ◽  
Z. A. Kanarskaya ◽  
A. V. Scherbakov ◽  
E. N. Scherbakova ◽  
...  

Xylanase is an enzyme that hydrolyses β-1,4 bonds in plant xylan. This enzyme is applied in the bioconversion of agro-industrial waste for xylooligosaccharide hydrolysate production to improve digestibility and nutrition value of animal feed, food processing, the utilisation and faster decomposition of crop debris in soil, as well as in cellulose bleaching and other industries. The current trend focuses on using renewable resources, such as agricultural waste, as substitutes for expensive purified xylan in producer screening and xylanase synthesis. This work aimed to determine the impact of Paenibacillus mucilaginosus cultivation conditions on the xylanase production yield. Rice bran ferment lysate along with birch and beech timber xylans were used as a carbon source. Temperature, medium pH, pH correction factors, inoculant incubation time, carbon and nitrogen sources and concentrations were the studied criteria of xylanase biosynthesis and growth in bacteria P. ucilaginosus strain 560. We show that the xylanase biosynthesis and cultivation in P. mucilaginosus strain 560 are more practical and cost-effective with the use of a rice bran ferment lysate-based nutrient medium. Inductors contained in the rice bran ferment lysate improve the xylanase biosynthesis. Calcium ions also facilitate biosynthesis in the studied strain. Cultivation recommendations are: carbon source concentration in medium 0.5% of total reducing substances content; 0.2% carbamide as optimal nitrogen source; calcium hydroxide as an agent for medium pH correction to 6.0±0.2; cultivation temperature 30±1 °С. Under the specified conditions, cultivation of P. mucilaginosus does not require inoculate preprocessing, and a maximal xylanase activity in stationary culture reaches 20 U/mL.

2016 ◽  
Vol 6 (01) ◽  
Author(s):  
Erika Erika ◽  
Rochmah Agustrina ◽  
Sumardi Sumardi ◽  
Mulyono Mulyono

Xylan is a carbon source in growth medium of extracellular xylanase producing bacteria. The purpose of this study was to get the optimum medium for the growth of Bacillus sp. in producing the xylanase. The factors consist of production time, carbon, and nitrogen source, as well as simple sugars. Addition carbon source used was delignified sugarcane bagasse, rice hulls, and corn cobs with different concentrations (0.25%; 0.5%; 0.75%; and 1% w/v) . Ammonium chloride, ammonium sulfate, and sodium nitrate with different concentrations (0.08%; 0.17%; 0.26%; and 0.35% w/v) were used as a source of nitrogen, while the simple sugar used was glucose, lactose, sucrose, and xylose. The results showed that the optimum culture media of Bacillus sp. to produce xylanase is media with 0.25% natural starch from the corn cob xylan as a carbon source, 0.26% ammonium chloride as a source of nitrogen, 0.0625 grams of sugar xylose, at pH 6, incubation temperature of 40°C, and 12 hours production time. In that media, xylanase activity was 0.2 U/mL.Keywords: agricultural waste, medium optimization, xylanase, Bacillus sp.   ABSTRAKXilan merupakan sumber karbon pada media pertumbuhan bakteri penghasil enzim ekstraseluler xilanase. Tujuan penelitian ini adalah mendapatkan media optimum untuk pertumbuhan Bacillus sp. dalam memproduksi xilanase. Perlakuan percobaan terdiri dari waktu produksi, sumber karbon, sumber nitrogen, dan penambahan gula sederhana. Sumber karbon yang digunakan adalah bagas tebu, sekam padi, dan tongkol jagung dengan variasi konsentrasi 0,25%; 0,5%; 0,75%; dan 1% (b/v) . Amonium klorida, amonium sulfat, dan natrium nitrat dengan variasi konsentrasi 0,08%; 0,17%; 0,26%; dan 0,35% (b/v) digunakan sebagai sumber nitrogen, sedangkan gula sederhana yang digunakan adalah glukosa, laktosa, sukrosa, dan xilosa masing-masing sebanyak 0,0625 b/v. Hasil percobaan menunjukkan bahwa media optimum pertumbuhan Bacillus sp. untuk produksi xilanase adalah media dengan 0,25% tepung xilan dari tongkol jagung sebagai sumber karbon, 0,26% amonium klorida sebagai sumber nitrogen, 0,0625 gram gula xilosa, pada pH media 6, suhu inkubasi 40°C, serta waktu produksi 12 jam. Dalam media tersebut, aktivitas xilanase yang dihasilkan sebesar 0,2 U/mL.Kata kunci : limbah pertanian, optimasi media, xilanase, Bacillus sp. 


2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Guo-Qiang Guan ◽  
Peng-Xiang Zhao ◽  
Jin Zhao ◽  
Mei-Juan Wang ◽  
Shu-Hao Huo ◽  
...  

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.


2021 ◽  
Author(s):  
Romain Bouchat ◽  
Frédéric Vélard ◽  
Sandrine Audonnet ◽  
Damien Rioult ◽  
Frank Delvigne ◽  
...  

Abstract Background: The microbial production of hemicellulasic cocktails is still a challenge for the sector of biorefineries and agro-waste valorization. In this work, the production of hemicellulolytic enzymes by Thermobacillus xylanilyticus has been considered. This microorganism is of interest since it is able to produce an original set of thermostable hemicellulolytic enzymes, and notably a xylanase GH11, Tx-xyn11. However, cell-to-cell heterogeneities impairs the production capability of the whole microbial population.Results: Sequential cultivations of the strain on xylan as a carbon source has been considered in order to highlight and better understand this cell-to-cell heterogeneity. Successive cultivations pointed out a fast decrease of xylanase activity (loss of ~75%) after 23.5 generations. Accordingly, the expression of the Tx-xyn11 gene decreased drastically and followed the same trend as the xylanase activity. Flow cytometry analyses pointed out that two subpopulations, differing at the level of their light scattering properties, were potentially involved in this progressive loss of enzymatic activities. Interestingly, upon successive cultivations on xylan, the subpopulation exhibiting low forward scatter (FSC) signal. Additionally, the evolution of the ratio between the two subpopulations was correlated to the decrease in xylanase activity. Cell sorting and direct observation of the sorted subpopulations revealed that the low-FSC subpopulation was not sporulating, whereas the high-FSC subpopulation contained cells at the onset of the sporulation stage. Serial cultivations on glucose, followed by the addition of a xylan pulse led to a ~1.5-fold to ~15-fold improvement of xylanase, depending on the moment for pulse addition, , suggesting that alternating cultivation conditions could lead to an efficient population management strategy for the production of xylanase. Conclusions: Taken altogether, the data from this study point out that a cheating behaviour is responsible for the progressive reduction in xylanase activity during serial cultivations of T. xylanilyticus. Alternating cultivation condition between glucose and xylan could be used as an efficient strategy for promoting population stability and higher enzymatic productivity from this bacterium.


2011 ◽  
Vol 2011 ◽  
pp. 1-9 ◽  
Author(s):  
Abbas Akhavan Sepahy ◽  
Shokoofeh Ghazi ◽  
Maryam Akhavan Sepahy

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.


2019 ◽  
Vol 19 (2) ◽  
pp. 470 ◽  
Author(s):  
Siti Nor Amira Rosli ◽  
Rohaida Che Man ◽  
Nasratun Masngut

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.


2009 ◽  
Vol 76 (3) ◽  
pp. 359-364
Author(s):  
S.M. Tauk-Tornisiel ◽  
M.C. Vallejo ◽  
J.C. Govone

ABSTRACT Six Penicillium strains were isolated from soil at a depth of 0 15 cm in the Juréia-Itatins Ecology Station (JIES), in the São Paulo State, Brazil. They were evaluated for xylanase production under different temperatures and carbon sources. The best carbon source and temperature were first determined in an automated Bioscreen C system, verifying the growth of microorganisms. Liquid media containing tap water with 2% carbohydrate and/or 1% nitrogen sources were used. Afterwards, Penicillium citrinum, P. fellutanum, P. rugulosum and P. decumbens were cultivated in 250 mL Erlenmeyer flasks with 50 mL of culture medium containing tap water sole 2% carbon source (fructose, glucose, mannitol, sucrose or xylose) and 1% yeast extract as a nitrogen source at pH 5.0 and 28o C, with agitation of 150 rpm for 72 hours. These same strains, except P. decumbens, and P. purpurogenum were cultivated in solid substrate with wheat bran under the same environmental conditions to study the potential of xylanase activity. Maximum xylanase activity was observed in cultures with wheat bran, without the addition of any other carbon source, using inocula containing 1 x 107 spores.mL-1 (28o C, pH 5.0, 72 h). It can be concluded that P. fellutanum and P. citrinumare a good xylanase producers under the conditions of 28º C. The results of xylanase activity were 54% less at 28º C in liquid cultures media cultures than in solid substrate.


2019 ◽  
Vol 50 (3) ◽  
Author(s):  
Al-Badran & Al-Shamary

 Seventeen local isolates of Bacillus were isolated from soil to produce extracellular xylanase under submerged fermentation process by using xylan as carbon sole source. All isolates were subjected  to quantitative scanning to select the most efficient one. The highest activity of xylanase (2680u/ml) was obtained from isolate Bacillus sp RS1. The isolate identified by 16SrRNA gene sequence of Bacillus subtilis  ( accuracy of 99%)which was matched with sequence of Bacillus subtilis VBN25 that recorded in Genebank under the Accession Number of MG027675.1.Extracted xylan from agricultural waste by acidic method(papyrus, sun flower stalks, Ibaa Wheat type, Furat wheat type and Abo Ghraib wheat type)were used as the substrate for xylanase production from Bacillus. The results  showed that the papyrus gave the highest amount of xylan (187.6 µg/ml) as compared with that of the sun flower stalks, Ibaa Wheat type, Furat wheat type and Abo Ghraib wheat type(161.3, 161.6, 157.6, 157.2) µ g/ml respectively. The results indicated that the highest  xylanase activity was 2800 u/ml produced by Bacillus subtilus when Papyrus xylan was used.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Marta Matuszewska ◽  
Tomasz Maciąg ◽  
Magdalena Rajewska ◽  
Aldona Wierzbicka ◽  
Sylwia Jafra

AbstractPseudomonas donghuensis P482 is a tomato rhizosphere isolate with the ability to inhibit growth of bacterial and fungal plant pathogens. Herein, we analysed the impact of the carbon source on the antibacterial activity of P482 and expression of the selected genes of three genomic regions in the P482 genome. These regions are involved in the synthesis of pyoverdine, 7-hydroxytropolone (7-HT) and an unknown compound (“cluster 17”) and are responsible for the antimicrobial activity of P482. We showed that the P482 mutants, defective in these regions, show variations and contrasting patterns of growth inhibition of the target pathogen under given nutritional conditions (with glucose or glycerol as a carbon source). We also selected and validated the reference genes for gene expression studies in P. donghuensis P482. Amongst ten candidate genes, we found gyrB, rpoD and mrdA the most stably expressed. Using selected reference genes in RT-qPCR, we assessed the expression of the genes of interest under minimal medium conditions with glucose or glycerol as carbon sources. Glycerol was shown to negatively affect the expression of genes necessary for 7-HT synthesis. The significance of this finding in the light of the role of nutrient (carbon) availability in biological plant protection is discussed.


2021 ◽  
Vol 22 (8) ◽  
pp. 4214
Author(s):  
Gautam Anand ◽  
Meirav Leibman-Markus ◽  
Dorin Elkabetz ◽  
Maya Bar

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.


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