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

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.

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.


2017 ◽  
Vol 30 (11) ◽  
pp. 886-895 ◽  
Author(s):  
Maria Chiara Paccanaro ◽  
Luca Sella ◽  
Carla Castiglioni ◽  
Francesca Giacomello ◽  
Ana Lilia Martínez-Rocha ◽  
...  

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.


2020 ◽  
Vol 14 (1) ◽  
pp. 5
Author(s):  
SIPRIYADI SIPRIYADI ◽  
ARIS TRI WAHYUDI ◽  
MAGGY THENAWIDJAYA SUHARTONO ◽  
ANJA MERYANDINI

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.


2019 ◽  
Vol 35 (6) ◽  
pp. 57-66
Author(s):  
L.N. Borshchevskaya ◽  
A.N. Kalinina ◽  
S.P. Sineoky ◽  
M.D. Kashirskaya

Effect of overexpression of the HAC1 genes from Pichia pastoris and Saccharomyces cerevisiae on the production of heterologous enzymes, Escherichia coli phytase and Paenibacillus brasilensis xylanase, in P. pastoris cells has been studied. Codon composition of the phytase and xylanase encoding genes was optimized, and the genes were expressed in P. pastoris under the control of AOX1 promoter. The obtained multi-copy strains produced in vitro 927 U/mL phytase and 1,401 U/ml xylanase activity. Overexpression of the HAC1 gene from P. pastoris was shown to increase the phytase and xylanase production by 46% and 41%, respectively. Overexpression of HAC1 from S. cerevisiae increased the phytase production by 28% and xylanase by 20%. Data obtained could be helpful in the construction of industrial enzyme-producing strains based on P. pastoris. phytase, xylanase, Нас1р transcription activator, UPR, Pichia pastoris, Saccharomyces cerevisiae The work was carried out using Multipurpose Scientific Installation All-Russian Collection of Industrial Microorganisms National Bioresource Center, NRC «Kurchatov Institute» -GosNIIgenetika. The authors are grateful to A.V. Nikulin (Sintol LLC, Russia) for the assistance in the real-time PCR experiments. The work was financially supported by the Ministry of Education and Science of the Russian Federation (Unique Identifiers of the Projects are RFMEFI57917X0145 and RFMEFI60717X0180).


1970 ◽  
Vol 45 (2) ◽  
pp. 151-154
Author(s):  
Apurba Lal Ray ◽  
Tahsina Rahim

Neurospora crassa, a non-pathogenic filamentous fungus produces hydrolytic enzymes (cellulase, xylanase, etc.) and is capable to convert lignocellulosic materials (cellulose, xylan) into simple sugars. Carboxymethylcellulase (CMCase) and xylanase activities were evaluated using 4 morphological mutants of N. crassa induced with leaf extract of Abroma augusta L in comparison to the wild type (Ema). Enzymatic activity was expressed as International Unit (IU). It was found that N. crassa mycelia secrete cellulases (CMCase) and xylanases to the culture supernatant. Wild N. crassa (Ema) showed highest cellulase activity (0.066 IU) and mutant ro 197 showed lowest (0.013 IU) activity. In case of xylanase, wild showed highest (0.313 IU) and mutant ro 197 showed lowest (0.088 IU) activity. These enzymatic assays revealed that wild and mutants of N. crassa possess detectable cellulase and xylanase activity. Key words: Mutants; Neurospora crassa; Enzyme activitiesDOI: 10.3329/bjsir.v45i2.5713Bangladesh J. Sci. Ind. Res. 45(2), 151-154, 2010


Fermentation ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 59
Author(s):  
Timothy J. Tse ◽  
Daniel J. Wiens ◽  
Jianheng Shen ◽  
Aaron D. Beattie ◽  
Martin J. T. Reaney

As barley and oat production have recently increased in Canada, it has become prudent to investigate these cereal crops as potential feedstocks for alcoholic fermentation. Ethanol and other coproduct yields can vary substantially among fermented feedstocks, which currently consist primarily of wheat and corn. In this study, the liquified mash of milled grains from 28 barley (hulled and hull-less) and 12 oat cultivars were fermented with Saccharomyces cerevisiae to determine concentrations of fermentation products (ethanol, isopropanol, acetic acid, lactic acid, succinic acid, α-glycerylphosphorylcholine (α-GPC), and glycerol). On average, the fermentation of barley produced significantly higher amounts of ethanol, isopropanol, acetic acid, succinic acid, α-GPC, and glycerol than that of oats. The best performing barley cultivars were able to produce up to 78.48 g/L (CDC Clear) ethanol and 1.81 g/L α-GPC (CDC Cowboy). Furthermore, the presence of milled hulls did not impact ethanol yield amongst barley cultivars. Due to its superior ethanol yield compared to oats, barley is a suitable feedstock for ethanol production. In addition, the accumulation of α-GPC could add considerable value to the fermentation of these cereal crops.


Fermentation ◽  
2019 ◽  
Vol 5 (1) ◽  
pp. 16 ◽  
Author(s):  
Luis Huezo ◽  
Ajay Shah ◽  
Frederick Michel

Previous studies have shown that pretreatment of corn slurries using ultrasound improves starch release and ethanol yield during biofuel production. However, studies on its effects on the mass transfer of substrates and products during fermentation have shown that it can have both beneficial and inhibitory effects. In this study, the effects of ultrasound on mass transfer limitations during fermentation were examined. Calculation of the external and intraparticle observable moduli under a range of conditions indicate that no external or intraparticle mass transfer limitations should exist for the mass transfer of glucose, ethanol, or carbon dioxide. Fermentations of glucose to ethanol using Saccharomyces cerevisiae were conducted at different ultrasound intensities to examine its effects on glucose uptake, ethanol production, and yeast population and viability. Four treatments were compared: direct ultrasound at intensities of 23 and 32 W/L, indirect ultrasound (1.4 W/L), and no-ultrasound. Direct and indirect ultrasound had negative effects on yeast performance and viability, and reduced the rates of glucose uptake and ethanol production. These results indicate that ultrasound during fermentation, at the levels applied, is inhibitory and not expected to improve mass transfer limitations.


2013 ◽  
Vol 34 (3) ◽  
pp. 253-267 ◽  
Author(s):  
Mauro Tropeano ◽  
Susana Vázquez ◽  
Silvia Coria ◽  
Adrián Turjanski ◽  
Daniel Cicero ◽  
...  

AbstractCold−adapted marine bacteria producing extracellular hydrolytic enzymes are important for their industrial application and play a key role in degradation of particulate organic matter in their natural environment. In this work, members of a previously−obtained protease−producing bacterial collection isolated from different marine sources from Potter Cove (King George Island, South Shetlands) were taxonomically identified and screened for their ability to produce other economically relevant enzymes. Eighty−eight proteolytic bacterial isolates were grouped into 25 phylotypes based on their Amplified Ribosomal DNA Restriction Analysis profiles. The sequencing of the 16S rRNA genes from representative isolates of the phylotypes showed that the predominant culturable protease−producing bacteria belonged to the class Gammaproteobacteria and were affiliated to the genera Pseudomonas, Shewanella, Colwellia, and Pseudoalteromonas, the latter being the predominant group (64% of isolates). In addition, members of the classes Actinobacteria, Bacilli and Flavobacteria were found. Among the 88 isolates screened we detected producers of amylases (21), pectinases (67), cellulases (53), CM−cellulases (68), xylanases (55) and agarases (57). More than 85% of the isolates showed at least one of the extracellular enzymatic activities tested, with some of them producing up to six extracellular enzymes. Our results confirmed that using selective conditions to isolate producers of one extracellular enzyme activity increases the probability of recovering bacteria that will also produce additional extracellular enzymes. This finding establishes a starting point for future programs oriented to the prospecting for biomolecules in Antarctica.


Fermentation ◽  
2018 ◽  
Vol 4 (3) ◽  
pp. 59 ◽  
Author(s):  
Tingting Liu ◽  
Shuangcheng Huang ◽  
Anli Geng

Cost-effective production of cellulosic ethanol requires robust microorganisms for rapid co-fermentation of glucose and xylose. This study aims to develop a recombinant diploid xylose-fermenting Saccharomyces cerevisiae strain for efficient conversion of lignocellulosic biomass sugars to ethanol. Episomal plasmids harboring codon-optimized Piromyces sp. E2 xylose isomerase (PirXylA) and Orpinomyces sp. ukk1 xylose (OrpXylA) genes were constructed and transformed into S. cerevisiae. The strain harboring plasmids with tandem PirXylA was favorable for xylose utilization when xylose was used as the sole carbon source, while the strain harboring plasmids with tandem OrpXylA was beneficial for glucose and xylose cofermentation. PirXylA and OrpXylA genes were also individually integrated into the genome of yeast strains in multiple copies. Such integration was beneficial for xylose alcoholic fermentation. The respiration-deficient strain carrying episomal or integrated OrpXylA genes exhibited the best performance for glucose and xylose co-fermentation. This was partly attributed to the high expression levels and activities of xylose isomerase. Mating a respiration-efficient strain carrying the integrated PirXylA gene with a respiration-deficient strain harboring integrated OrpXylA generated a diploid recombinant xylose-fermenting yeast strain STXQ with enhanced cell growth and xylose fermentation. Co-fermentation of 162 g L−1 glucose and 95 g L−1 xylose generated 120.6 g L−1 ethanol in 23 h, with sugar conversion higher than 99%, ethanol yield of 0.47 g g−1, and ethanol productivity of 5.26 g L−1·h−1.


2015 ◽  
Vol 9 (7) ◽  
pp. 8 ◽  
Author(s):  
Tri Widjaja ◽  
Ali Altway ◽  
Arief Widjaja ◽  
Umi Rofiqah ◽  
Rr Whiny Hardiyati Erlian

One form of economic development efforts for waste utilization in rural communities is to utilize stem sorghum to produce food grade ethanol. Sorghum stem juice with 150 g/L of sugar concentration was fermented using conventional batch process and cell immobilization continuous process with K-carrageenan as a supporting matrix. The microorganism used was Mutated Zymomonas Mobilis to be compared with a mixture of Saccharomyces Cerevisiae and Pichia Stipitis, and a mixture of Mutated Zymomonas Mobilis and Pichia Stipitis. Ethanol in the broth, result of fermentation process, was separated in packed distillation column. Distilate of the column, still contain water and other impurities, was flown into molecular sieve for dehydration and activated carbon adsorption column to remove the other impurities to meet food grade ethanol specification. The packing used in distillation process was steel wool. For batch fermentation, the fermentation using a combination of Saccharomyces Cerevisiae and Pichia Stipitis produced the best ethanol with 12.07% of concentration, where the yield and the productivity were 63.49%, and 1.06 g/L.h, respectively. And for continuous fermentation, the best ethanol with 9.02% of concentration, where the yield and the productivity were 47.42% and 174.27 g/L.h, respectively, is obtained from fermentation using a combination of Saccharomyces Cerevisiae and Pichia Stipitis also. Fermentation using combination microorganism of Saccharomyces Cerevisiae and Pichia Stipitis produced higher concentration of ethanol, yield, and productivity than other microorganisms. Distillation, molecular sieve dehydration and adsorption process is quite successful in generating sufficient levels of ethanol with relatively low amount of impurities.


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