scholarly journals Consolidated bioprocessing of lignocellulose for production of glucaric acid by an artificial microbial consortium

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Chaofeng Li ◽  
Xiaofeng Lin ◽  
Xing Ling ◽  
Shuo Li ◽  
Hao Fang

Abstract Background The biomanufacturing of d-glucaric acid has attracted increasing interest because it is one of the top value-added chemicals produced from biomass. Saccharomyces cerevisiae is regarded as an excellent host for d-glucaric acid production. Results The opi1 gene was knocked out because of its negative regulation on myo-inositol synthesis, which is the limiting step of d-glucaric acid production by S. cerevisiae. We then constructed the biosynthesis pathway of d-glucaric acid in S. cerevisiae INVSc1 opi1Δ and obtained two engineered strains, LGA-1 and LGA-C, producing record-breaking titers of d-glucaric acid: 9.53 ± 0.46 g/L and 11.21 ± 0.63 g/L d-glucaric acid from 30 g/L glucose and 10.8 g/L myo-inositol in fed-batch fermentation mode, respectively. However, LGA-1 was preferable because of its genetic stability and its superior performance in practical applications. There have been no reports on d-glucaric acid production from lignocellulose. Therefore, the biorefinery processes, including separated hydrolysis and fermentation (SHF), simultaneous saccharification and fermentation (SSF) and consolidated bioprocessing (CBP) were investigated and compared. CBP using an artificial microbial consortium composed of Trichoderma reesei (T. reesei) Rut-C30 and S. cerevisiae LGA-1 was found to have relatively high d-glucaric acid titers and yields after 7 d of fermentation, 0.54 ± 0.12 g/L d-glucaric acid from 15 g/L Avicel and 0.45 ± 0.06 g/L d-glucaric acid from 15 g/L steam-exploded corn stover (SECS), respectively. In an attempt to design the microbial consortium for more efficient CBP, the team consisting of T. reesei Rut-C30 and S. cerevisiae LGA-1 was found to be the best, with excellent work distribution and collaboration. Conclusions Two engineered S. cerevisiae strains, LGA-1 and LGA-C, with high titers of d-glucaric acid were obtained. This indicated that S. cerevisiae INVSc1 is an excellent host for d-glucaric acid production. Lignocellulose is a preferable substrate over myo-inositol. SHF, SSF, and CBP were studied, and CBP using an artificial microbial consortium of T. reesei Rut-C30 and S. cerevisiae LGA-1 was found to be promising because of its relatively high titer and yield. T. reesei Rut-C30 and S. cerevisiae LGA-1were proven to be the best teammates for CBP. Further work should be done to improve the efficiency of this microbial consortium for d-glucaric acid production from lignocellulose.

2021 ◽  
Author(s):  
Chaofeng Li ◽  
Xiaofeng Lin ◽  
Xing Ling ◽  
Shuo Li ◽  
Hao Fang

Abstract Background: The biomanufacturing of D-glucaric acid has been attracted increasing interest because it is one of the top value-added chemicals produced from biomass. Saccharomyces cerevisiae is regarded as an excellent host for D-glucaric acid production. Results: The opi1 gene was knocked out because of its negative regulation on myo-inositol synthesis, which is the limiting step of D-glucaric acid production by S. cerevisiae. Then we constructed the biosynthetic pathway of D-glucaric acid in S. cerevisiae INVSc1 opi1Δ and obtained two engineered strains, LGA-1 and LGA-C, producing record breaking titers of D-glucaric acid, 9.53 ± 0.46 g/L and 11.21 ± 0.63 g/L D-glucaric acid from 30 g/L glucose and 10.8 g/L myo-inositol in the mode of fed-batch fermentation, respectively. However, LGA-1 was more preferable because of the genetic stability and the outperformance in applications. So far, there have been no reports on D-glucaric acid production from lignocellulose. Therefore, the biorefinery processes including separated hydrolysis and fermentation (SHF), simultaneous saccharification and fermentation (SSF) and consolidated bioprocessing (CBP) were investigated and compared. CBP by an artificial microbial consortium composed of Trichoderma reesei Rut-C30 and S. cerevisiae LGA-1 was found to have relatively high D-glucaric acid titers and yields after 7 d fermentation, 0.54 ± 0.12 g/L D-glucaric acid from 15 g/L Avicel, and 0.45 ± 0.06 g/L D-glucaric acid from 15 g/L steam exploded corn stover (SECS), respectively. In attempts to design the microbial consortium for more efficient CBP the team consisted of the two members, T. reesei Rut-C30 and S. cerevisiae LGA-1, was found to be the best with excellent work distribution and collaboration.Conclusions: Two engineered S. cerevisiae strains, LGA-1 and LGA-C, with high titers of D-glucaric acid were obtained. This indicates that S. cerevisiae INVSc1 was an excellent host. Lignocellulose is a more preferable substrate than myo-inositol. SHF, SSF and CBP were studied and CBP by an artificial microbial consortium of T. reesei Rut-C30 and S. cerevisiae LGA-1 was found to be promising because of the relatively high titer and yield. T. reesei Rut-C30 and S. cerevisiae LGA-1were proved to be the best teammates for CBP. Further work should be done to improve the efficiency of this microbial consortium for D-glucaric acid production from lignocellulose.


2020 ◽  
Author(s):  
Chaofeng Li ◽  
Xiaofeng Lin ◽  
Xing Ling ◽  
Shuo Li ◽  
Hao Fang

AbstractThe biomanufacturing of D-glucaric acid has been attracted increasing interest and the industrial yeast Saccharomyces cerevisiae is regarded as an excellent host for D-glucaric acid production. Here we constructed the biosynthetic pathway of D-glucaric acid in S. cerevisiae INVSc1 whose opi1 was knocked out and obtained two engineered strains, LGA-1 and LGA-C, producing record breaking titers of D-glucaric acid, 9.53 ± 0.46 g/L and 11.21 ± 0.63 g/L D-glucaric acid from 30 g/L glucose and 10.8 g/L myo-inositol in the mode of fed-batch fermentation, respectively. Due to the genetic stability and the outperformance in subsequent applications, however, LGA-1 was a preferable strain. As one of the top chemicals from biomass, there have been no reports on D-glucaric acid production from lignocellulose, which is the most abundant renewable on earth. Therefore, the biorefinery processes of lignocellulose for D-glucaric acid production including separated hydrolysis and fermentation (SHF), simultaneous saccharification and fermentation (SSF) and consolidated bioprocessing (CBP) were investigated in this work and CBP by an artificial microbial consortium composed of Trichoderma reesei Rut-C30 and S. cerevisiae LGA-1 was found to have relatively high D-glucaric acid titers and yields after 7 d fermentation, 0.54 ± 0.12 g/L D-glucaric acid from 15 g/L Avicel, and 0.45 ± 0.06 g/L D-glucaric acid from 15 g/L steam exploded corn stover (SECS), respectively. In attempts to design the microbial consortium for more efficient CBP the team consisted of the two members, T. reesei Rut-C30 and S. cerevisiae LGA-1, was found to be the best with excellent work distribution and collaboration. This desirable and promising approach for direction production of D-glucaric acid from lignocellulose deserves extensive and in-depth research.


2020 ◽  
Author(s):  
Chaofeng Li ◽  
Xiaofeng Lin ◽  
Xing Ling ◽  
Shuo Li ◽  
Hao Fang

Abstract Background The biomanufacturing of D-glucaric acid has been attracted increasing interest and the industrial yeast Saccharomyces cerevisiae is regarded as an excellent host for D-glucaric acid production. Results Here we constructed the biosynthetic pathway of D-glucaric acid in S. cerevisiae INVSc1 whose opi1 was knocked out and obtained two engineered strains, LGA-1 and LGA-C, producing record breaking titers of D-glucaric acid, 9.53 ± 0.46 g/L and 11.21 ± 0.63 g/L D-glucaric acid from 30 g/L glucose and 10.8 g/L myo -inositol in the mode of fed-batch fermentation, respectively. Due to the genetic stability and the outperformance in subsequent applications, however, LGA-1 was a preferable strain. As one of the top chemicals from biomass, there have been no reports on D-glucaric acid production from lignocellulose, which is the most abundant renewable on earth. Therefore, the biorefinery processes of lignocellulose for D-glucaric acid production including separated hydrolysis and fermentation (SHF), simultaneous saccharification and fermentation (SSF) and consolidated bioprocessing (CBP) were investigated in this work and CBP by an artificial microbial consortium composed of Trichoderma reesei Rut-C30 and S. cerevisiae LGA-1 was found to have relatively high D-glucaric acid titers and yields after 7 d fermentation, 0.54 ± 0.12 g/L D-glucaric acid from 15 g/L Avicel, and 0.45 ± 0.06 g/L D-glucaric acid from 15 g/L steam exploded corn stover (SECS), respectively. Conclusions In attempts to design the microbial consortium for more efficient CBP the team consisted of the two members, T. reesei Rut-C30 and S. cerevisiae LGA-1, was found to be the best with excellent work distribution and collaboration. This desirable and promising approach for direction production of D-glucaric acid from lignocellulose deserves extensive and in-depth research.


Author(s):  
Tai D. Nguyen ◽  
Ronald Gronsky ◽  
Jeffrey B. Kortright

Nanometer period Ru/C multilayers are one of the prime candidates for normal incident reflecting mirrors at wavelengths < 10 nm. Superior performance, which requires uniform layers and smooth interfaces, and high stability of the layered structure under thermal loadings are some of the demands in practical applications. Previous studies however show that the Ru layers in the 2 nm period Ru/C multilayer agglomerate upon moderate annealing, and the layered structure is no longer retained. This agglomeration and crystallization of the Ru layers upon annealing to form almost spherical crystallites is a result of the reduction of surface or interfacial energy from die amorphous high energy non-equilibrium state of the as-prepared sample dirough diffusive arrangements of the atoms. Proposed models for mechanism of thin film agglomeration include one analogous to Rayleigh instability, and grain boundary grooving in polycrystalline films. These models however are not necessarily appropriate to explain for the agglomeration in the sub-nanometer amorphous Ru layers in Ru/C multilayers. The Ru-C phase diagram shows a wide miscible gap, which indicates the preference of phase separation between these two materials and provides an additional driving force for agglomeration. In this paper, we study the evolution of the microstructures and layered structure via in-situ Transmission Electron Microscopy (TEM), and attempt to determine the order of occurence of agglomeration and crystallization in the Ru layers by observing the diffraction patterns.


2019 ◽  
Vol 6 (1) ◽  
Author(s):  
Hui-Hui Su ◽  
Fei Peng ◽  
Pei Xu ◽  
Xiao-Ling Wu ◽  
Min-Hua Zong ◽  
...  

Abstract Background Glucaric acid, one of the aldaric acids, has been declared a “top value-added chemical from biomass”, and is especially important in the food and pharmaceutical industries. Biocatalytic production of glucaric acid from glucuronic acid is more environmentally friendly, efficient and economical than chemical synthesis. Uronate dehydrogenases (UDHs) are the key enzymes for the preparation of glucaric acid in this way, but the poor thermostability and low activity of UDH limit its industrial application. Therefore, improving the thermostability and activity of UDH, for example by semi-rational design, is a major research goal. Results In the present work, three UDHs were obtained from different Agrobacterium tumefaciens strains. The three UDHs have an approximate molecular weight of 32 kDa and all contain typically conserved UDH motifs. All three UDHs showed optimal activity within a pH range of 6.0–8.5 and at a temperature of 30 °C, but the UDH from A. tumefaciens (At) LBA4404 had a better catalytic efficiency than the other two UDHs (800 vs 600 and 530 s−1 mM−1). To further boost the catalytic performance of the UDH from AtLBA4404, site-directed mutagenesis based on semi-rational design was carried out. An A39P/H99Y/H234K triple mutant showed a 400-fold improvement in half-life at 59 °C, a 5 °C improvement in $$ {\text{T}}_{ 5 0}^{ 1 0} $$ T 50 10 value and a 2.5-fold improvement in specific activity at 30 °C compared to wild-type UDH. Conclusions In this study, we successfully obtained a triple mutant (A39P/H99Y/H234K) with simultaneously enhanced activity and thermostability, which provides a novel alternative for the industrial production of glucaric acid from glucuronic acid.


Catalysts ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 887
Author(s):  
Rujeeluk Khumho ◽  
Satit Yousatit ◽  
Chawalit Ngamcharussrivichai

5-Hydroxymethylfurfural (HMF) is one of the most important lignocellulosic biomass-derived platform molecules for production of renewable fuel additives, liquid hydrocarbon fuels, and value-added chemicals. The present work developed niobium oxides (Nb2O5) supported on mesoporous carbon/silica nanocomposite (MCS), as novel solid base catalyst for synthesis of HMF via one-pot glucose conversion in a biphasic solvent. The MCS material was prepared via carbonization using natural rubber dispersed in hexagonal mesoporous silica (HMS) as a precursor. The Nb2O5 supported on MCS (Nb/MCS) catalyst with an niobium (Nb) loading amount of 10 wt.% (10-Nb/MCS) was characterized by high dispersion, and so tiny crystallites of Nb2O5, on the MCS surface, good textural properties, and the presence of Bronsted and Lewis acid sites with weak-to-medium strength. By varying the Nb loading amount, the crystallite size of Nb2O5 and molar ratio of Bronsted/Lewis acidity could be tuned. When compared to the pure silica HMS-supported Nb catalyst, the Nb/MCS material showed a superior glucose conversion and HMF yield. The highest HMF yield of 57.5% was achieved at 93.2% glucose conversion when using 10-Nb/MCS as catalyst (5 wt.% loading with respect to the mass of glucose) at 190 °C for 1 h. Furthermore, 10-Nb/MCS had excellent catalytic stability, being reused in the reaction for five consecutive cycles during which both the glucose conversion and HMF yield were insignificantly changed. Its superior performance was ascribed to the suitable ratio of Brønsted/Lewis acid sites, and the hydrophobic properties generated from the carbon moieties dispersed in the MCS nanocomposite.


Coatings ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 558
Author(s):  
Emilia-Adela Salca ◽  
Tomasz Krystofiak ◽  
Barbara Lis ◽  
Salim Hiziroglu

The objective of this study was to evaluate the glossiness of black alder wood (Alnus glutinosa L.) samples coated with two varnish types as a function of exposure to dry heat and artificial aging. The chemical resistance of the coated samples to cold liquids was also evaluated. Based on the findings in this work, it appears that the varnish types and their structural differences influenced the overall glossiness of the coated samples. The UV varnish exhibited higher gloss values than those coated with the water-borne product within the range of silky gloss and silky matte grades. The heat exposure influenced the surface glossiness of the UV-coated samples more than the samples coated with water-borne varnish. The overall gloss values of the samples decreased with the exposure time to artificial aging, resulting in no layer cracks. The cold household liquids left less visible traces on the surfaces and alcohol was found to be the strongest agent. This study could have practical applications in the furniture industry to produce value-added furniture units according to their specific conditions of indoor use.


2021 ◽  
Vol 4 ◽  
Author(s):  
Debomitra Dey ◽  
Jana K. Richter ◽  
Pichmony Ek ◽  
Bon-Jae Gu ◽  
Girish M. Ganjyal

The processing of agricultural products into value-added food products yields numerous by-products or waste streams such as pomace (fruit and vegetable processing), hull/bran (grain milling), meal/cake (oil extraction), bagasse (sugar processing), brewer's spent grain (brewing), cottonseed meal (cotton processing), among others. In the past, significant work in exploring the possibility of the utilization of these by-products has been performed. Most by-products are highly nutritious and can be excellent low-cost sources of dietary fiber, proteins, and bioactive compounds such as polyphenols, antioxidants, and vitamins. The amount of energy utilized for the disposal of these materials is far less than the energy required for the purification of these materials for valorization. Thus, in many cases, these materials go to waste or landfill. Studies have been conducted to incorporate the by-products into different foods in order to promote their utilization and tackle their environmental impacts. Extrusion processing can be an excellent avenue for the utilization of these by-products in foods. Extrusion is a widely used thermo-mechanical process due to its versatility, flexibility, high production rate, low cost, and energy efficiency. Extruded products such as direct-expanded products, breakfast cereals, and pasta have been developed by researchers using agricultural by-products. The different by-products have a wide range of characteristics in terms of chemical composition and functional properties, affecting the final products in extrusion processing. For the practical applications of these by-products in extrusion, it is crucial to understand their impacts on the qualities of raw material blends and extruded products. This review summarizes the general differences in the properties of food by-products from different sources (proximate compositions, physicochemical properties, and functional properties) and how these properties and the extrusion processing conditions influence the product characteristics. The discussion of the by-product properties and their impacts on the extrudates and their nutritional profile can be useful for food manufacturers and researchers to expand their applications. The gaps in the literature have been highlighted for further research and better utilization of by-products with extrusion processing.


2019 ◽  
Author(s):  
RAN DU ◽  
YUE HU ◽  
René Hübner ◽  
Jan-Ole Joswig ◽  
Xuelin Fan ◽  
...  

<div>Noble metal foams (NMFs) are a new class of functional porous materials featuring properties of both noble metals and monolithic porous materials, providing impressive prospects in catalysis, bio-sensing, plasmonic technologies, etc...Among reported synthetic methods to date, the sol-gel approach manifests overwhelming advantages for versatile synthesis of controlled nanostructured NMFs under mild condition. However, limited gelation methods and insufficient understanding of the underlying mechanism retards structure/composition manipulation of NMFs, hampering ondemand designing for practical applications. Herein highly tunable NMFs are fabricated at room temperature by activating specific-ion effects and regulating ion-nanoparticle interactions, affording various single/alloy NMFs with adjustable compositions (Au, Ag, Pd, Pt), ligament sizes (3.1~142.0 nm), and special morphologies. Their superior performance in programmable self-propulsion devices and electrocatalytic alcohol oxidation are demonstrated. This study provides not only a conceptually new route to fabricate and manipulate functional NMFs, but also an overall picture in understanding the gelation mechanism. It may pave the way for on-target designing versatile NMFs for various applications.</div>


2021 ◽  
Vol 28 ◽  
Author(s):  
Yuyang Xue ◽  
Xiucai Ye ◽  
Lesong Wei ◽  
Xin Zhang ◽  
Tetsuya Sakurai ◽  
...  

: With its superior performance, the Transformer model, which is based on the 'Encoder-Decoder' paradigm, has become the mainstream in natural language processing. On the other hand, bioinformatics has embraced machine learning and made great progress in drug design and protein property prediction. Cell-penetrating peptides (CPPs) are one kind of permeable protein that is convenient as a kind of 'postman' in drug penetration tasks. However, a small number of CPPs have been discovered by research, let alone practical applications in drug permeability. Therefore, correctly identifying the CPPs has opened up a new way to take macromolecules into cells without other potentially harmful materials in the drug. Most of the previous work only uses trivial machine learning techniques and hand-crafted features to construct a simple classifier. In CPPFormer, we learn from the idea of implementing the attention structure of Transformer, rebuilding the network based on the characteristics of CPPs according to its short length, and using an automatic feature extractor with a few manual engineered features to co-direct the predicted results. Compared to all previous methods and other classic text classification models, the empirical result has shown that our proposed deep model-based method has achieved the best performance of 92.16% accuracy in the CPP924 dataset and has passed various index tests.


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