scholarly journals Dissecting cellobiose metabolic pathway and its application in biorefinery through consolidated bioprocessing in Myceliophthora thermophila

2019 ◽  
Vol 6 (1) ◽  
Author(s):  
Jingen Li ◽  
Shuying Gu ◽  
Zhen Zhao ◽  
Bingchen Chen ◽  
Qian Liu ◽  
...  
Keyword(s):  
Malic Acid ◽  
Plant Biomass ◽  
Consolidated Bioprocessing ◽  
Cellulase Activity ◽  
Industrial Applications ◽  
Efficient Production ◽  
Myceliophthora Thermophila ◽  
Final Strain ◽  
Cellobiose Metabolism ◽  
Cost Efficient

Abstract Background Lignocellulosic biomass has long been recognized as a potential sustainable source for industrial applications. The costs associated with conversion of plant biomass to fermentable sugar represent a significant barrier to the production of cost-competitive biochemicals. Consolidated bioprocessing (CBP) is considered a potential breakthrough for achieving cost-efficient production of biomass-based fuels and commodity chemicals. During the degradation of cellulose, cellobiose (major end-product of cellulase activity) is catabolized by hydrolytic and phosphorolytic pathways in cellulolytic organisms. However, the details of the two intracellular cellobiose metabolism pathways in cellulolytic fungi remain to be uncovered. Results Using the engineered malic acid production fungal strain JG207, we demonstrated that the hydrolytic pathway by β-glucosidase and the phosphorolytic pathway by phosphorylase are both used for intracellular cellobiose metabolism in Myceliophthora thermophila, and the yield of malic acid can benefit from the energy advantages of phosphorolytic cleavage. There were obvious differences in regulation of the two cellobiose catabolic pathways depending on whether M. thermophila JG207 was grown on cellobiose or Avicel. Disruption of Mtcpp in strain JG207 led to decreased production of malic acid under cellobiose conditions, while expression levels of all three intracellular β-glucosidase genes were significantly up-regulated to rescue the impairment of the phosphorolytic pathway under Avicel conditions. When the flux of the hydrolytic pathway was reduced, we found that β-glucosidase encoded by bgl1 was the dominant enzyme in the hydrolytic pathway and deletion of bgl1 resulted in significant enhancement of protein secretion but reduction of malate production. Combining comprehensive manipulation of both cellobiose utilization pathways and enhancement of cellobiose uptake by overexpression of a cellobiose transporter, the final strain JG412Δbgl2Δbgl3 produced up to 101.2 g/L and 77.4 g/L malic acid from cellobiose and Avicel, respectively, which corresponded to respective yields of 1.35 g/g and 1.03 g/g, representing significant improvement over the starting strain JG207. Conclusions This is the first report of detailed investigation of intracellular cellobiose catabolism in cellulolytic fungus M. thermophila. These results provide insights that can be applied to industrial fungi for production of biofuels and biochemicals from cellobiose and cellulose.

scholarly journals Coordination of consolidated bioprocessing technology and carbon dioxide fixation to produce malic acid directly from plant biomass in Myceliophthora thermophila

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Jingen Li ◽  
Bingchen Chen ◽  
Shuying Gu ◽  
Zhen Zhao ◽  
Qian Liu ◽  
...  
Keyword(s):  
Malic Acid ◽  
Plant Biomass ◽  
Consolidated Bioprocessing ◽  
Average Rate ◽  
Value Added ◽  
Cost Effective ◽  
Myceliophthora Thermophila ◽  
Carbon Efficiency ◽  
Bulk Chemicals ◽  
Promising Strategy

Abstract Background Consolidated bioprocessing (CBP) technique is a promising strategy for biorefinery construction, producing bulk chemicals directly from plant biomass without extra hydrolysis steps. Fixing and channeling CO2 into carbon metabolism for increased carbon efficiency in producing value-added compounds is another strategy for cost-effective bio-manufacturing. It has not been reported whether these two strategies can be combined in one microbial platform. Results In this study, using the cellulolytic thermophilic fungus Myceliophthora thermophila, we designed and constructed a novel biorefinery system DMCC (Direct microbial conversion of biomass with CO2 fixation) through incorporating two CO2 fixation modules, PYC module and Calvin–Benson–Bassham (CBB) pathway. Harboring the both modules, the average rate of fixing and channeling 13CO2 into malic acid in strain CP51 achieved 44.4, 90.7, and 80.7 mg/L/h, on xylose, glucose, and cellulose, respectively. The corresponding titers of malic acid were up to 42.1, 70.4, and 70.1 g/L, respectively, representing the increases of 40%, 10%, and 7%, respectively, compared to the parental strain possessing only PYC module. The DMCC system was further improved by enhancing the pentose uptake ability. Using raw plant biomass as the feedstock, yield of malic acid produced by the DMCC system was up to 0.53 g/g, with 13C content of 0.44 mol/mol malic acid, suggesting DMCC system can produce 1 t of malic acid from 1.89 t of biomass and fix 0.14 t CO2 accordingly. Conclusions This study designed and constructed a novel biorefinery system named DMCC, which can convert raw plant biomass and CO2 into organic acid efficiently, presenting a promising strategy for cost-effective production of value-added compounds in biorefinery. The DMCC system is one of great options for realization of carbon neutral economy.

scholarly journals Dynamic consolidated bioprocessing for innovative lab-scale production of bacterial alkaline phosphatase from Bacillus paralicheniformis strain APSO

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Soad A. Abdelgalil ◽  
Nadia A. Soliman ◽  
Gaber A. Abo-Zaid ◽  
Yasser R. Abdel-Fattah
Keyword(s):  
Alkaline Phosphatase ◽  
Consolidated Bioprocessing ◽  
Black Liquor ◽  
Batch Cultivation ◽  
Efficient Production ◽  
Scale Production ◽  
Market Demand ◽  
Stirred Tank Bioreactor ◽  
Bacterial Alkaline Phosphatase ◽  
Bacillus Paralicheniformis

AbstractTo meet the present and forecasted market demand, bacterial alkaline phosphatase (ALP) production must be increased through innovative and efficient production strategies. Using sugarcane molasses and biogenic apatite as low-cost and easily available raw materials, this work demonstrates the scalability of ALP production from a newfound Bacillus paralicheniformis strain APSO isolated from a black liquor sample. Mathematical experimental designs including sequential Plackett–Burman followed by rotatable central composite designs were employed to select and optimize the concentrations of the statistically significant media components, which were determined to be molasses, (NH4)2NO3, and KCl. Batch cultivation in a 7-L stirred-tank bioreactor under uncontrolled pH conditions using the optimized medium resulted in a significant increase in both the volumetric and specific productivities of ALP; the alkaline phosphatase throughput 6650.9 U L−1, and µ = 0.0943 h−1; respectively, were obtained after 8 h that, ameliorated more than 20.96, 70.12 and 94 folds compared to basal media, PBD, and RCCD; respectively. However, neither the increased cell growth nor enhanced productivity of ALP was present under the pH-controlled batch cultivation. Overall, this work presents novel strategies for the statistical optimization and scaling up of bacterial ALP production using biogenic apatite.

scholarly journals Screening and directed evolution of transporters to improve xylodextrin utilization in the yeast Saccharomyces cerevisiae

2017 ◽  
Author(s):  
Chenlu Zhang ◽  
Ligia Acosta-Sampson ◽  
Vivian Yaci Yu ◽  
Jamie H. D. Cate
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Directed Evolution ◽  
Plant Biomass ◽  
Carbon Sources ◽  
Industrial Applications ◽  
Yeast Saccharomyces Cerevisiae ◽  
Cellulosic Biofuel ◽  
Economic Production ◽  
Report Analysis ◽  
Selection Medium

AbstractThe economic production of cellulosic biofuel requires efficient and full utilization of all abundant carbohydrates naturally released from plant biomass by enzyme cocktails. Recently, we reconstituted the Neurospora crassa xylodextrin transport and consumption system in Saccharomyces cerevisiae, enabling growth of yeast on xylodextrins aerobically. However, the consumption rate of xylodextrin requires improvement for industrial applications, including consumption in anaerobic conditions. As a first step in this improvement, we report analysis of orthologues of the N. crassa transporters CDT-1 and CDT-2. Transporter ST16 from Trichoderma virens enables faster aerobic growth of S. cerevisiae on xylodextrins compared to CDT-2. ST16 is a xylodextrin-specific transporter, and the xylobiose transport activity of ST16 is not inhibited by cellobiose. Other transporters identified in the screen also enable growth on xylodextrins including xylotriose. Taken together, these results indicate that multiple transporters might prove useful to improve xylodextrin utilization in S. cerevisiae. Efforts to use directed evolution to improve ST16 from a chromosomally-integrated copy were not successful, due to background growth of yeast on other carbon sources present in the selection medium. Future experiments will require increasing the baseline growth rate of the yeast population on xylodextrins, to ensure that the selective pressure exerted on xylodextrin transport can lead to isolation of improved xylodextrin transporters.

scholarly journals perekonomian indonesa

2019 ◽  
Author(s):  
Pricyllia
Keyword(s):  
Sugar Industry ◽  
Raw Material ◽  
Efficient Production ◽  
Internal Conflicts ◽  
Beverage Industry ◽  
Self Sufficiency ◽  
Food And Beverage Industry ◽  
Food And Beverage ◽  
Cost Efficient ◽  
The Cost

The 1930s was the heyday of the Indonesian sugarindustry that is capable of exporting to manycountries and has become the country an exporterof sugar to two after Cuba, but the situation isreversed since 1967 in which Indonesia wouldimport sugar from Brazil, India, and Thailand inorder to meet the needs raw material consumptionand food and beverage industry. The resultsshowed that the cost of sugar production isvery uneconomical because of inefficiency thatstretches from the cultivation to processing inthe factory so difficult to obtain profit margins.Target and beyond sugar self-sufficiency can notbe achieved because highly regulated, there is nosynergy and tends to conflicts of interests amongministries or agencies, and internal conflicts oftenoccur between the sugar mill and the disharmonybetween sugarcane farmers by the sugar millofficials.In the competition of the sugar industry aremore stringent, in the era of the Asean EconomicCommunity, it means the level of efficiency of sugarfactories in the country need special attention,the same thing also applies to industrial users ofsugar, and sugar as a raw material componentscontribute to the creation of products food andbeverage efficient so as to compete with similarproducts from other countries. Observing howthe intense competition in the Asean EconomicCommunity is based on free trade, the yield ofsugar that’s a cost efficient production is veryimportant and urgent at this time, including workto improve the welfare of farmers.

scholarly journals Evolution of a Biomass-Fermenting Bacterium To Resist Lignin Phenolics

2017 ◽  
Vol 83 (11) ◽  
Author(s):  
Tristan Cerisy ◽  
Tiffany Souterre ◽  
Ismael Torres-Romero ◽  
Magali Boutard ◽  
Ivan Dubois ◽  
...  
Keyword(s):  
Ferulic Acid ◽  
Plant Biomass ◽  
Consolidated Bioprocessing ◽  
Gene Dosage ◽  
Content Type ◽  
Chemical Inhibitors ◽  
Growth Selection ◽  
Clostridium Phytofermentans

ABSTRACT Increasing the resistance of plant-fermenting bacteria to lignocellulosic inhibitors is useful to understand microbial adaptation and to develop candidate strains for consolidated bioprocessing. Here, we study and improve inhibitor resistance in Clostridium phytofermentans (also called Lachnoclostridium phytofermentans), a model anaerobe that ferments lignocellulosic biomass. We survey the resistance of this bacterium to a panel of biomass inhibitors and then evolve strains that grow in increasing concentrations of the lignin phenolic, ferulic acid, by automated, long-term growth selection in an anaerobic GM3 automat. Ultimately, strains resist multiple inhibitors and grow robustly at the solubility limit of ferulate while retaining the ability to ferment cellulose. We analyze genome-wide transcription patterns during ferulate stress and genomic variants that arose along the ferulate growth selection, revealing how cells adapt to inhibitors through changes in gene dosage and regulation, membrane fatty acid structure, and the surface layer. Collectively, this study demonstrates an automated framework for in vivo directed evolution of anaerobes and gives insight into the genetic mechanisms by which bacteria survive exposure to chemical inhibitors. IMPORTANCE Fermentation of plant biomass is a key part of carbon cycling in diverse ecosystems. Further, industrial biomass fermentation may provide a renewable alternative to fossil fuels. Plants are primarily composed of lignocellulose, a matrix of polysaccharides and polyphenolic lignin. Thus, when microorganisms degrade lignocellulose to access sugars, they also release phenolic and acidic inhibitors. Here, we study how the plant-fermenting bacterium Clostridium phytofermentans resists plant inhibitors using the lignin phenolic, ferulic acid. We examine how the cell responds to abrupt ferulate stress by measuring changes in gene expression. We evolve increasingly resistant strains by automated, long-term cultivation at progressively higher ferulate concentrations and sequence their genomes to identify mutations associated with acquired ferulate resistance. Our study develops an inhibitor-resistant bacterium that ferments cellulose and provides insights into genomic evolution to resist chemical inhibitors.

Targeted analysis of phenolic compounds by LC-MS v1

2021 ◽  
Author(s):  
Bashar Amer ◽  
Ramu Kakumanu ◽  
yangtian not provided ◽  
Aymerick Eudes ◽  
Edward EK Baidoo
Keyword(s):  
Phenolic Compounds ◽  
Plant Biomass ◽  
Reversed Phase ◽  
Industrial Applications ◽  
Liquid Chromatography Mass Spectrometry ◽  
Reversed Phase Liquid Chromatography ◽  
Sample Matrix ◽  
Targeted Analysis ◽  
Challenges And Opportunities ◽  
Phase Liquid

Cell-wall-bound (CWB) aromatics such as ferulate and p-coumarate play important physiological roles in plant development and response to stresses. Their presence also poses some challenges and opportunities during processing of plant biomass in various agro-industrial applications. To this end, we have developed a robust high-throughput reversed-phase liquid chromatography mass spectrometry method for quantifying CWB phenolic compounds. The method showed excellent linearity (R2 = ≥0.999) and intraday retention time repeatability (≤ 0.31 %RSD) for ferulate and p-coumarate. The limits of detection and quantitation for these analytes were ≤ 39 nM and 130 nM, respectively. Furthermore, there was very little effect of the CWB sample matrix on the retention times of the analytes and analyte percent recoveries from the CWB sample matrix was ≥83.91%.

A novel Swollenin from Talaromyces leycettanus JCM12802 exhibits cellulase activity and enhanced cellulase hydrolysis towards various substrates

2020 ◽  
Author(s):  
Honghai Zhang ◽  
Yuan Wang ◽  
Roman Brunecky ◽  
Bin Yao ◽  
Xiangming Xie ◽  
...  
Keyword(s):  
Synergistic Effect ◽  
Functional Diversity ◽  
Specific Activity ◽  
Biomass Conversion ◽  
Hydrolytic Enzymes ◽  
Cellulase Activity ◽  
Industrial Applications ◽  
Maximum Activity ◽  
Carbohydrate Binding ◽  
Cell Wall Polysaccharides

Abstract Background Swollenins are present in some fungal species involved in the biodegradation of cellulosic substrates. They appear to promote a rearrangement in the network of non-covalent interactions between the cell wall polysaccharides, thus making it more accessible for degradation by hydrolytic enzymes. Here, we have reported a detailed characterization of a recombinant swollenin with respect to its disruptive activity on cellulosic substrates and synergistic effect with cellulases. Results In the present study, a novel swollenin gene Tlswo consisting of an open reading frame encoding 503 amino acids was identified from Talaromyces leycettanus JCM12802 and successfully expressed in Trichoderma reesei and Pichia pastoris. Similar to other fungal swollenins, TlSWO contained a N-terminal family 1 carbohydrate binding module (CBM1) followed by a Ser/Thr rich linker connected to expansin-like domain which includes a family 45 endoglucanase-like domain and group-2 grass pollen allergen domain. TlSWO demonstrated disruptive activity on Avicel and displayed a high synergistic effect with cellobiohydrolases, enhancing its hydrolytic performance up to 132%. The activity of TlSWO on various substrates and biomass was also examined. It was shown that TlSWO could release reducing sugars from lichenan, barley β-glucan, carboxymethyl cellulose sodium (CMC-Na) and laminarin. The specific activity of TlSWO towards the substates above is 9.0 ± 0.100 U/mg, 8.9 ± 0.100U/mg, 2.3 ± 0.002 U/mg and 0.79 ± 0.002 U/mg respectively. Moreover, TlSWO exhibits maximum activity at pH 4.0 and 50 ℃. Conclusion This study reported on a novel swollenin with highly efficient for biomass conversion. It also reveals the functional diversity of swollenin with activity on various substrates. Although the exact mechanism of swollenin catalytic action activity still remains unknown, the functional diversity of TlSWO makes it a good candidate for industrial applications.

Optimization of Culture Conditions for Cold-Active Cellulase Production by Penicillium cordubense D28 Using Response Surface Methodology

2011 ◽  
Vol 183-185 ◽  
pp. 994-998
Author(s):  
Shuo Dong ◽  
Nai Yu Chi ◽  
Qing Fang Zhang
Keyword(s):  
Response Surface Methodology ◽  
Response Surface ◽  
Cellulase Activity ◽  
Cellulase Production ◽  
Culture Conditions ◽  
Batch Reactors ◽  
Corn Meal ◽  
Cold Active ◽  
Initial Ph ◽  
Cost Efficient

The design of an optimum and cost-efficient medium for production of cold-active cellulase by Penicillium cordubense D28 was attempted by using response surface methodology (RSM). Based on the Plackett–Burman design, corn meal, (NH4)2SO4 and branc were selected as the most critical nutrient. Subsequently, they were investigated by the Box-Behnken design. Results showed that the maximum cold-active cellulase activity of 110.4U/mL was predicted when the concentration of corn meal, (NH4)2SO4 and branc were 21.97 g/L, 2.39 g/L and 14.99 g/L, respectively. The results were further verified by triplicate experiments. The batch reactors were operated under an optimized condition of the respective corn meal, (NH4)2SO4 and branc concentration of 22 g/L , 2.4 g/L and 15 g/L , the initial pH of 6.0 and experimental temperature of 20 ± 1°C. Without further pH adjustment, the maximum cold-active cellulase activity of 109.8 U/mL was obtained based on the optimized medium with further verified the practicability of this optimum strategy.

scholarly journals A Fast Method for Predicting the Mechanical Properties of Precipitation-Hardenable Aluminum Alloys

Metals ◽  
2019 ◽  
Vol 9 (2) ◽  
pp. 147 ◽  
Author(s):  
Anastasiya Toenjes ◽  
Axel von Hehl
Keyword(s):  
Mechanical Properties ◽  
Heat Treatment ◽  
Aluminum Alloys ◽  
Method Development ◽  
Prediction Models ◽  
Industrial Applications ◽  
Fast Method ◽  
Suitable Heat Treatment ◽  
Cost Efficient ◽  
The Individual

Most heat treatment simulations of precipitation-hardenable aluminum alloys are incomplete or restricted to sub-steps of the process chain. In general, the studies addressing the heat treatment of aluminum components have only provided a qualitative guidance of heat treatment, which does not match the heat treatment that is necessary for specific parts with specific requirements. Thus, a quick and accurate simulation of the whole heat treatment process would hold great economic benefit for industrial applications in predicting suitable heat treatment processes that are able to meet the required mechanical properties of proposed novel aluminum components. In this paper, the development of a time and cost efficient method for generating such prediction models is presented by means of an example aluminum alloy EN AW-6082. During the process sub-steps of solution annealing, quenching and aging, the time-temperature correlations connected to the precipitation-hardening conditions were analyzed. The precision of the prediction model depends on the size of the material database, which should be able to be adjusted to the individual requirements of the simulation user. In order to obtain the greatest time and cost efficiency in generating such a model, a specific experimental design was developed. The results of the method development are presented and discussed.

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