Bioresources and Bioprocessing

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ISSN / EISSN : 2197-4365 / 2197-4365
Published by: Springer Nature (10.1186)
Total articles ≅ 449
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Dongdong Chang, Cong Wang, Fabrice Ndayisenga, Zhisheng Yu
Published: 22 October 2021
Bioresources and Bioprocessing, Volume 8, pp 1-14; https://doi.org/10.1186/s40643-021-00459-2

Abstract:
Levoglucosan is a promising sugar present in the lignocellulose pyrolysis bio-oil, which is a renewable and environment-friendly source for various value-added productions. Although many microbial catalysts have been engineered to produce biofuels and chemicals from levoglucosan, the demerits that these biocatalysts can only utilize pure levoglucosan while inhibited by the inhibitors co-existing with levoglucosan in the bio-oil have greatly limited the industrial-scale application of these biocatalysts in lignocellulose biorefinery. In this study, the previously engineered Escherichia coli LGE2 was evolved for enhanced inhibitor tolerance using long-term adaptive evolution under the stress of multiple inhibitors and finally, a stable mutant E. coli-H was obtained after ~ 374 generations’ evolution. In the bio-oil media with an extremely acidic pH of 3.1, E. coli-H with high inhibitor tolerance exhibited remarkable levoglucosan consumption and ethanol production abilities comparable to the control, while the growth of the non-evolved strain was completely blocked even when the pH was adjusted to 7.0. Finally, 8.4 g/L ethanol was achieved by E. coli-H in the undetoxified bio-oil media with ~ 2.0% (w/v) levoglucosan, reaching 82% of the theoretical yield. Whole-genome re-sequencing to monitor the acquisition of mutations identified 4 new mutations within the globally regulatory genes rssB, yqhA, and basR, and the − 10 box of the putative promoter of yqhD-dgkA operon. Especially, yqhA was the first time to be revealed as a gene responsible for inhibitor tolerance. The mutations were all responsible for improved fitness, while basR mutation greatly contributed to the fitness improvement of E. coli-H. This study, for the first time, generated an inhibitor-tolerant levoglucosan-utilizing strain that could produce cost-effective bioethanol from the toxic bio-oil without detoxification process, and provided important experimental evidence and valuable genetic/proteinic information for the development of other robust microbial platforms involved in lignocellulose biorefining processes.
Pengye Guo, Yuanchan Luo, Ju Wu,
Published: 22 October 2021
Bioresources and Bioprocessing, Volume 8, pp 1-17; https://doi.org/10.1186/s40643-021-00458-3

Abstract:
Due to the increasing environmental pollution of un-degradable plastics and the consumption of non-renewable resources, more attention has been attracted by new bio-degradable/based polymers produced from renewable resources. Polylactic acid (PLA) is one of the most representative bio-based materials, with obvious advantages and disadvantages, and has a wide range of applications in industry, medicine, and research. By copolymerizing to make up for its deficiencies, the obtained copolymers have more excellent properties. The development of a one-step microbial metabolism production process of the lactate (LA)-based copolymers overcomes the inherent shortcomings in the traditional chemical synthesis process. The most common lactate-based copolymer is poly(lactate-co-3-hydroxybutyrate) [P(LA-co-3HB)], within which the difference of LA monomer fraction will cause the change in the material properties. It is necessary to regulate LA monomer fraction by appropriate methods. Based on synthetic biology and systems metabolic engineering, this review mainly focus on how did the different production strategies (such as enzyme engineering, fermentation engineering, etc.) of P(LA-co-3HB) optimize the chassis cells to efficiently produce it. In addition, the metabolic engineering strategies of some other lactate-based copolymers are also introduced in this article. These studies would facilitate to expand the application fields of the corresponding materials.
Su Yan, Yan Xu, Xiao-Wei Yu
Published: 22 October 2021
Bioresources and Bioprocessing, Volume 8, pp 1-15; https://doi.org/10.1186/s40643-021-00461-8

Abstract:
The filamentous fungus Trichoderma reesei has been widely used for cellulase production that has extensive applications in green and sustainable development. Increasing costs and depletion of fossil fuels provoke the demand for hyper-cellulase production in this cellulolytic fungus. To better manipulate T. reesei for enhanced cellulase production and to lower the cost for large-scale fermentation, it is wise to have a comprehensive understanding of the crucial factors and complicated biological network of cellulase production that could provide new perspectives for further exploration and modification. In this review, we summarize recent progress and give an overview of the cellular process of cellulase production in T. reesei, including the carbon source-dependent cellulase induction, complicated transcriptional regulation network, and efficient protein assembly and trafficking. Among that, the key factors involved in cellulase production were emphasized, shedding light on potential perspectives for further engineering.
Yimei Xi, Song Xue, Xupeng Cao, Zhanyou Chi, Jinghan Wang
Published: 21 October 2021
Bioresources and Bioprocessing, Volume 8, pp 1-11; https://doi.org/10.1186/s40643-021-00457-4

Abstract:
Accumulation of β-carotene in Dunaliella salina is highly dependent on light exposure intensity and duration, but quantitative analysis on photon numbers received per cell for triggering β-carotene accumulation is not available so far. In this study, experiment results showed that significant β-carotene accumulation occurred after at least 8 h illumination at 400 µmol photons·m−2·s−1. To quantify the average number of photons received per cell, correlations of light attenuation with light path, biomass concentration, and β-carotene content were, respectively, established using both Lambert–Beer and Cornet models, and the latter provided better simulation. Using Cornet model, average number of photons received per cell (APRPC) was calculated and proposed as a parameter for β-carotene accumulation, and constant APRPC was maintained by adjusting average irradiance based on cell concentration and carotenoids content changes during the whole induction period. It was found that once APRPC reached 0.7 µmol photons cell−1, β-carotene accumulation was triggered, and it was saturated at 9.9 µmol photons cell−1. This study showed that APRPC can be used as an important parameter to precisely simulate and control β-carotene production by D. salina. Graphic Abstract
Zi-Fu Ni, Pei Xu, Min-Hua Zong,
Published: 19 October 2021
Bioresources and Bioprocessing, Volume 8, pp 1-9; https://doi.org/10.1186/s40643-021-00456-5

Abstract:
Enzymatic asymmetric amination addition is seen as a promising approach for synthesizing amine derivatives, especially unnatural amino acids, which are valuable precursors to fine chemicals and drugs. Despite the broad substrate spectrum of methylaspartate lyase (MAL), some bulky substrates, such as caffeic acid, cannot be effectively accepted. Herein, we report a group of variants structurally derived from Escherichia coli O157:H7 MAL (EcMAL). A combined mutagenesis strategy was used to simultaneously redesign the key residues of the entrance tunnel and binding pocket to explore the possibility of accepting bulky substrates with potential application to chiral drug synthesis. Libraries of residues capable of lining the active center of EcMAL were then constructed and screened by an effective activity solid-phase color screening method using tyrosinase as a cascade catalyst system. Activity assays and molecular dynamics studies of the resultant variants showed that the substrate specificity of EcMAL was modified by adjusting the polarity of the binding pocket and the degree of flexibility of the entrance tunnel. Compared to M3, the optimal variant M8 was obtained with a 15-fold increase in catalytic activity. This structure-based protein engineering of EcMAL can be used to open new application directions or to develop practical multi-enzymatic processes for the production of various useful compounds.
Jianwei Feng, Shenyuan Xu, Ruirui Feng, Andrey Kovalevsky, Xia Zhang, Dongyang Liu,
Published: 17 October 2021
Bioresources and Bioprocessing, Volume 8, pp 1-12; https://doi.org/10.1186/s40643-021-00449-4

Abstract:
β-1,3-glucanase can specifically hydrolyze glucans to oligosaccharides and has potential applications in biotechnology. We used the metatranscriptomic technology to discover a thermophilic β-1,3-glucanase from compost. The phylogenetic study shows that it belongs to the family 16 glycoside hydrolase (GH16) and is most homologous with an enzyme from Streptomyces sioyaensis, an actinobacterium. It has the activity of 146.9 U/mg in the optimal reaction condition (75 °C and pH 5.5). Its catalytic domain was crystallized and diffracted to 1.14 Å resolution. The crystal structure shows a sandwich-like β-jelly-roll fold with two disulfide bonds. After analyzing the occurring frequencies of these cysteine residues, we designed two mutants (C160G and C180I) to study the role of these disulfide bonds. Both mutants have decreased their optimal temperature from 75 to 70 °C, which indicate that the disulfide bonds are important to maintain thermostability. Interestingly, the activity of C160G has increased ~ 17% to reach 171.4 U/mg. We speculate that the increased activity of C160G mutant is due to increased dynamics near the active site. Our studies give a good example of balancing the rigidity and flexibility for enzyme activity, which is helpful for protein engineering.
Ying-Chen Yi, I-Tai Shih, Tzu-Hsuan Yu, Yen-Ju Lee,
Published: 13 October 2021
Bioresources and Bioprocessing, Volume 8, pp 1-18; https://doi.org/10.1186/s40643-021-00455-6

Abstract:
5-Aminolevulinic acid (5-ALA), a non-proteinogenic five-carbon amino acid, has received intensive attentions in medicine due to its approval by the US Food and Drug Administration (FDA) for cancer diagnosis and treatment as photodynamic therapy. As chemical synthesis of 5-ALA performed low yield, complicated processes, and high cost, biosynthesis of 5-ALA via C4 (also called Shemin pathway) and C5 pathway related to heme biosynthesis in microorganism equipped more advantages. In C4 pathway, 5-ALA is derived from condensation of succinyl-CoA and glycine by 5-aminolevulic acid synthase (ALAS) with pyridoxal phosphate (PLP) as co-factor in one-step biotransformation. The C5 pathway involves three enzymes comprising glutamyl-tRNA synthetase (GltX), glutamyl-tRNA reductase (HemA), and glutamate-1-semialdehyde aminotransferase (HemL) from α-ketoglutarate in TCA cycle to 5-ALA and heme. In this review, we describe the recent results of 5-ALA production from different genes and microorganisms via genetic and metabolic engineering approaches. The regulation of different chassis is fine-tuned by applying synthetic biology and boosts 5-ALA production eventually. The purification process, challenges, and opportunities of 5-ALA for industrial applications are also summarized.
Shadhin, , Raghavan Jayaraman, Danny Mann
Published: 13 October 2021
Bioresources and Bioprocessing, Volume 8, pp 1-12; https://doi.org/10.1186/s40643-021-00453-8

Abstract:
Vacuum-assisted resin transfer molding (VARTM), used in manufacturing medium to large-sized composites for transportation industries, requires non-woven mats. While non-woven glass mats used in these applications are optimized for resin impregnation and properties, such optimized mats for natural fibers are not available. In the current research, cattail fibers were extracted from plants (18–30% yield) using alkali retting and non-woven cattail fiber mat was manufactured. The extracted fibers exhibited a normal distribution in diameter (d avg. = 32.1 µm); the modulus and strength varied inversely with diameter, and their average values were 19.1 GPa and 172.3 MPa, respectively. The cattail fiber composites were manufactured using non-woven mats, Stypol polyester resin, VARTM pressure (101 kPa) and compression molding pressures (260 and 560 kPa) and tested. Out-of-plane permeability changed with the fiber volume fraction (V f) of the mats, which was influenced by areal density, thickness, and fiber packing in the mat. The cattail fibers reinforced the Stypol resin significantly. The modulus and the strength increased with consolidation pressures due to the increase in V f, with maximum values of 7.4 GPa and 48 MPa, respectively, demonstrating the utility of cattail fibers from waste biomass as reinforcements.
, Fana Mulugeta Hagos, Rongrong Chen, Hanxin Qian, Chengxing Mo, Jing Di, Xikun Gai, Ruiqin Yang, Genxing Pan, Shengdao Shan
Published: 9 October 2021
Bioresources and Bioprocessing, Volume 8, pp 1-15; https://doi.org/10.1186/s40643-021-00451-w

Abstract:
Hydrochar a carbon-rich material resulting from hydrothermal carbonization of biomass, has received substantial attention because of its potential application in various areas such as carbon sequestration, bioenergy production and environmental amelioration. A series of hydrochars were prepared by metal chloride-assisted hydrothermal carbonization of rice husk and characterized by elemental analysis, zeta potential, X-ray diffraction, Brunauer–Emmett–Teller measurements, Fourier transform infrared spectroscopy, thermogravimetric analysis, X-ray photoelectron spectroscopy and scanning electron microscopy. The results reveal that the prepared hydrochars have carbon contents ranging from 45.01 to 58.71%, BET specific areas between 13.23 and 45.97 m2/g, and rich O-containing functional groups on the surfaces. The metal chlorides added in the feedwater could improve the degree of carbonization and show significant effects on the physical, chemical and adsorption properties of the hydrochars. The adsorption of the selected organics on the hydrochars is a spontaneous and physisorption-dominated process. The hydrochars possess larger adsorption capacities for 2-naphthol than for berberine hydrochloride and Congo red, and the modeling maximum adsorption capacities of 2-naphthol are in the range of 170.1–2680 mg/g. The adsorption equilibrium could be accomplished in 10, 40 and 30 min for 2-naphthol, berberine hydrochloride and Congo red, respectively. These results suggest metal chloride-assisted hydrothermal carbonization a promising method for converting biomass waste into effective adsorbents for wastewater treatment.
Doan Van Thuoc, Nguyen Thi Chung,
Published: 8 October 2021
Bioresources and Bioprocessing, Volume 8, pp 1-11; https://doi.org/10.1186/s40643-021-00454-7

Abstract:
Rice straw is an important low-cost feedstock for bio-based economy. This report presents a study in which rice straw was used both as a source for isolation of bacteria producing the biodegradable polyester polyhydroxyalkanoate (PHA), as well as the carbon source for the production of the polymer by the isolated bacteria. Of the 100 bacterial isolates, seven were found to be positive for PHA production by Nile blue staining and were identified as Bacillus species by 16S rRNA gene sequence analysis. Three isolates showed 100% sequence identity to B. cereus, one to B. paranthracis, two with 99 and 100% identity to B. anthracis, while one was closely similar to B. thuringiensis. For use in PHA production, rice straw was subjected to mild alkaline pretreatment followed by enzymatic hydrolysis. Comparison of pretreatment by 2% sodium hydroxide, 2% calcium hydroxide and 20% aqueous ammonia, respectively, at different temperatures showed maximum weight loss with NaOH at 80 °C for 5 h, but ammonia for 15 h at 80 °C led to highest lignin removal of 63%. The ammonia-pretreated rice straw also led to highest release of total reducing sugar up to 92% on hydrolysis by a cocktail of cellulases and hemicellulases at 50 °C. Cultivation of the Bacillus isolates on the pretreated rice straw revealed highest PHA content of 59.3 and 46.4%, and PHA concentration of 2.96 and 2.51 g/L by Bacillus cereus VK92 and VK98, respectively.
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