Biotechnology for Biofuels and Bioproducts最新文献

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A high-throughput dual system to screen polyphosphate kinase mutants for efficient ATP regeneration in L-theanine biocatalysis. 在L-茶氨酸生物催化中筛选高效ATP再生的多磷酸激酶突变体的高通量双系统。
Biotechnology for Biofuels and Bioproducts Pub Date : 2023-08-03 DOI: 10.1186/s13068-023-02361-9
Hui Gao, Mengxuan Li, Qing Wang, Tingting Liu, Xian Zhang, Taowei Yang, Meijuan Xu, Zhiming Rao
{"title":"A high-throughput dual system to screen polyphosphate kinase mutants for efficient ATP regeneration in L-theanine biocatalysis.","authors":"Hui Gao, Mengxuan Li, Qing Wang, Tingting Liu, Xian Zhang, Taowei Yang, Meijuan Xu, Zhiming Rao","doi":"10.1186/s13068-023-02361-9","DOIUrl":"10.1186/s13068-023-02361-9","url":null,"abstract":"<p><p>ATP, an important cofactor, is involved in many biocatalytic reactions that require energy. Polyphosphate kinases (PPK) can provide energy for ATP-consuming reactions due to their cheap and readily available substrate polyphosphate. We determined the catalytic properties of PPK from different sources and found that PPK from Cytophaga hutchinsonii (ChPPK) had the best catalytic activity for the substrates ADP and polyP<sub>6</sub>. An extracellular-intracellular dual system was constructed to high-throughput screen for better catalytic activity of ChPPK mutants. Finally, the specific activity of ChPPK<sub>D82N-K103E</sub> mutant was increased by 4.3 times. Therefore, we focused on the production of L-theanine catalyzed by GMAS as a model of ATP regeneration. Supplying 150 mM ATP, GMAS enzyme could produce 16.8 ± 1.3 g/L L-theanine from 100 mM glutamate. When 5 mM ATP and 5 U/mL ChPPK<sub>D82N-K103E</sub> were added, the yield of L-theanine was 16.6 ± 0.79 g/L with the conversion rate of 95.6 ± 4.5% at 4 h. Subsequently, this system was scaled up to 200 mM and 400 mM glutamate, resulting in the yields of L-theanine for 32.3 ± 1.6 g/L and 62.7 ± 1.1 g/L, with the conversion rate of 92.8 ± 4.6% and 90.1 ± 1.6%, respectively. In addition, we also constructed an efficient ATP regeneration system from glutamate to glutamine, and 13.8 ± 0.2 g/L glutamine was obtained with the conversion rate of 94.4 ± 1.4% in 4 h after adding 6 U/ mL GS enzyme and 5 U/ mL ChPPK<sub>D82N-K103E</sub>, which further laid the foundation from glutamine to L-theanine catalyzed by GGT enzyme. This proved that giving the reaction an efficient ATP supply driven by the mutant enzyme enhanced the conversion rate of substrate to product and maximized the substrate value. This is a positively combination of high yield, high conversion rate and high economic value of enzyme catalysis. The mutant enzyme will further power the ATP-consuming biocatalytic reaction platform sustainably.</p>","PeriodicalId":9125,"journal":{"name":"Biotechnology for Biofuels and Bioproducts","volume":"16 1","pages":"122"},"PeriodicalIF":0.0,"publicationDate":"2023-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10401862/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10666625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 1
Alanine dehydrogenases from four different microorganisms: characterization and their application in L-alanine production. 四种不同微生物的丙氨酸脱氢酶:表征及其在l -丙氨酸生产中的应用。
Biotechnology for Biofuels and Bioproducts Pub Date : 2023-08-03 DOI: 10.1186/s13068-023-02373-5
Pengfei Gu, Qianqian Ma, Shuo Zhao, Qiang Li, Juan Gao
{"title":"Alanine dehydrogenases from four different microorganisms: characterization and their application in L-alanine production.","authors":"Pengfei Gu, Qianqian Ma, Shuo Zhao, Qiang Li, Juan Gao","doi":"10.1186/s13068-023-02373-5","DOIUrl":"10.1186/s13068-023-02373-5","url":null,"abstract":"<p><strong>Background: </strong>Alanine dehydrogenase (AlaDH) belongs to oxidoreductases, and it exists in several different bacteria species and plays a key role in microbial carbon and nitrogen metabolism, spore formation and photosynthesis. In addition, AlaDH can also be applied in biosynthesis of L-alanine from cheap carbon source, such as glucose.</p><p><strong>Results: </strong>To achieve a better performance of L-alanine accumulation, system evaluation and comparison of different AlaDH with potential application value are essential. In this study, enzymatic properties of AlaDH from Bacillus subtilis 168 (BsAlaDH), Bacillus cereus (BcAlaDH), Mycobacterium smegmatis MC<sup>2</sup> 155 (MsAlaDH) and Geobacillus stearothermophilus (GsAlaDH) were firstly carefully investigated. Four different AlaDHs have few similarities in optimum temperature and optimum pH, while they also exhibited significant differences in enzyme activity, substrate affinity and enzymatic reaction rate. The wild E. coli BL21 with these four AlaDHs could produce 7.19 g/L, 7.81 g/L, 6.39 g/L and 6.52 g/L of L-alanine from 20 g/L glucose, respectively. To further increase the L-alanine titer, competitive pathways for L-alanine synthesis were completely blocked in E. coli. The final strain M-6 could produce 80.46 g/L of L-alanine with a yield of 1.02 g/g glucose after 63 h fed-batch fermentation, representing the highest yield for microbial L-alanine production.</p><p><strong>Conclusions: </strong>Enzyme assay, biochemical characterization and structure analysis of BsAlaDH, BcAlaDH, MsAlaDH and GsAlaDH were carried out. In addition, application potential of these four AlaDHs in L-alanine productions were explored. The strategies here can be applied for developing L-alanine producing strains with high titers.</p>","PeriodicalId":9125,"journal":{"name":"Biotechnology for Biofuels and Bioproducts","volume":"16 1","pages":"123"},"PeriodicalIF":0.0,"publicationDate":"2023-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10401832/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9941191","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Unravelling and engineering an operon involved in the side-chain degradation of sterols in Mycolicibacterium neoaurum for the production of steroid synthons. 新金黄色分枝杆菌甾醇侧链降解以产生类固醇合成子的操纵子的研究。
Biotechnology for Biofuels and Bioproducts Pub Date : 2023-08-02 DOI: 10.1186/s13068-023-02376-2
Yun-Qiu Zhao, Yong-Jun Liu, Lu Song, Dingyan Yu, Kun Liu, Ke Liu, Bei Gao, Xin-Yi Tao, Liang-Bin Xiong, Feng-Qing Wang, Dong-Zhi Wei
{"title":"Unravelling and engineering an operon involved in the side-chain degradation of sterols in Mycolicibacterium neoaurum for the production of steroid synthons.","authors":"Yun-Qiu Zhao,&nbsp;Yong-Jun Liu,&nbsp;Lu Song,&nbsp;Dingyan Yu,&nbsp;Kun Liu,&nbsp;Ke Liu,&nbsp;Bei Gao,&nbsp;Xin-Yi Tao,&nbsp;Liang-Bin Xiong,&nbsp;Feng-Qing Wang,&nbsp;Dong-Zhi Wei","doi":"10.1186/s13068-023-02376-2","DOIUrl":"https://doi.org/10.1186/s13068-023-02376-2","url":null,"abstract":"<p><strong>Background: </strong>Harnessing engineered Mycolicibacteria to convert cheap phytosterols into valuable steroid synthons is a basic way in the industry for the production of steroid hormones. Thus, C-19 and C-22 steroids are the two main types of commercial synthons and the products of C17 side chain degradation of phytosterols. During the conversion process of sterols, C-19 and C-22 steroids are often produced together, although one may be the main product and the other a minor byproduct. This is a major drawback of the engineered Mycolicibacteria for industrial application, which could be attributed to the co-existence of androstene-4-ene-3,17-dione (AD) and 22-hydroxy-23,24-bisnorchol-4-ene-3-one (HBC) sub-pathways in the degradation of the sterol C17 side chain. Since the key mechanism underlying the HBC sub-pathway has not yet been clarified, the above shortcoming has not been resolved so far.</p><p><strong>Results: </strong>The key gene involved in the putative HBC sub-pathway was excavated from the genome of M. neoaurum by comparative genomic analysis. Interestingly, an aldolase- encoding gene, atf1, was identified to be responsible for the first reaction of the HBC sub-pathway, and it exists as a conserved operon along with a DUF35-type gene chsH4, a reductase gene chsE6, and a transcriptional regulation gene kstR3 in the genome. Subsequently, atf1 and chsH4 were identified as the key genes involved in the HBC sub-pathway. Therefore, an updated strategy was proposed to develop engineered C-19 or C-22 steroid-producing strains by simultaneously modifying the AD and HBC sub-pathways. Taking the development of 4-HBC and 9-OHAD-producing strains as examples, the improved 4-HBC-producing strain achieved a 20.7 g/L production titer with a 92.5% molar yield and a 56.4% reduction in byproducts, and the improved 9-OHAD producing strain achieved a 19.87 g/L production titer with a 94.6% molar yield and a 43.7% reduction in byproduct production.</p><p><strong>Conclusions: </strong>The excellent performances of these strains demonstrated that the primary operon involved in the HBC sub-pathway improves the industrial strains in the conversion of phytosterols to steroid synthons.</p>","PeriodicalId":9125,"journal":{"name":"Biotechnology for Biofuels and Bioproducts","volume":"16 1","pages":"121"},"PeriodicalIF":0.0,"publicationDate":"2023-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10398937/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9941394","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Correction: Secretion of collagenases by Saccharomyces cerevisiae for collagen degradation. 更正:酿酒酵母菌分泌胶原酶降解胶原。
Biotechnology for Biofuels and Bioproducts Pub Date : 2023-08-01 DOI: 10.1186/s13068-023-02372-6
Han Xiao, Xiufang Liu, Yunzi Feng, Lin Zheng, Mouming Zhao, Mingtao Huang
{"title":"Correction: Secretion of collagenases by Saccharomyces cerevisiae for collagen degradation.","authors":"Han Xiao,&nbsp;Xiufang Liu,&nbsp;Yunzi Feng,&nbsp;Lin Zheng,&nbsp;Mouming Zhao,&nbsp;Mingtao Huang","doi":"10.1186/s13068-023-02372-6","DOIUrl":"https://doi.org/10.1186/s13068-023-02372-6","url":null,"abstract":"","PeriodicalId":9125,"journal":{"name":"Biotechnology for Biofuels and Bioproducts","volume":"16 1","pages":"120"},"PeriodicalIF":0.0,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10394854/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9932731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Engineering Saccharomyces cerevisiae for improved biofilm formation and ethanol production in continuous fermentation. 在连续发酵中改进生物膜形成和乙醇生产的工程酿酒酵母。
Biotechnology for Biofuels and Bioproducts Pub Date : 2023-07-31 DOI: 10.1186/s13068-023-02356-6
Zhenyu Wang, Weikai Xu, Yixuan Gao, Mingwei Zha, Di Zhang, Xiwei Peng, Huifang Zhang, Cheng Wang, Chenchen Xu, Tingqiu Zhou, Dong Liu, Huanqing Niu, Qingguo Liu, Yong Chen, Chenjie Zhu, Ting Guo, Hanjie Ying
{"title":"Engineering Saccharomyces cerevisiae for improved biofilm formation and ethanol production in continuous fermentation.","authors":"Zhenyu Wang,&nbsp;Weikai Xu,&nbsp;Yixuan Gao,&nbsp;Mingwei Zha,&nbsp;Di Zhang,&nbsp;Xiwei Peng,&nbsp;Huifang Zhang,&nbsp;Cheng Wang,&nbsp;Chenchen Xu,&nbsp;Tingqiu Zhou,&nbsp;Dong Liu,&nbsp;Huanqing Niu,&nbsp;Qingguo Liu,&nbsp;Yong Chen,&nbsp;Chenjie Zhu,&nbsp;Ting Guo,&nbsp;Hanjie Ying","doi":"10.1186/s13068-023-02356-6","DOIUrl":"https://doi.org/10.1186/s13068-023-02356-6","url":null,"abstract":"<p><strong>Background: </strong>Biofilm-immobilized continuous fermentation has the potential to enhance cellular environmental tolerance, maintain cell activity and improve production efficiency.</p><p><strong>Results: </strong>In this study, different biofilm-forming genes (FLO5, FLO8 and FLO10) were integrated into the genome of S. cerevisiae for overexpression, while FLO5 and FLO10 gave the best results. The biofilm formation of the engineered strains 1308-FLO5 and 1308-FLO10 was improved by 31.3% and 58.7% compared to that of the WT strain, respectively. The counts of cells adhering onto the biofilm carrier were increased. Compared to free-cell fermentation, the average ethanol production of 1308, 1308-FLO5 and 1308-FLO10 was increased by 17.4%, 20.8% and 19.1% in the biofilm-immobilized continuous fermentation, respectively. Due to good adhering ability, the fermentation broth turbidity of 1308-FLO5 and 1308-FLO10 was decreased by 22.3% and 59.1% in the biofilm-immobilized fermentation, respectively. Subsequently, for biofilm-immobilized fermentation coupled with membrane separation, the engineered strain significantly reduced the pollution of cells onto the membrane and the membrane separation flux was increased by 36.3%.</p><p><strong>Conclusions: </strong>In conclusion, enhanced biofilm-forming capability of S. cerevisiae could offer multiple benefits in ethanol fermentation.</p>","PeriodicalId":9125,"journal":{"name":"Biotechnology for Biofuels and Bioproducts","volume":"16 1","pages":"119"},"PeriodicalIF":0.0,"publicationDate":"2023-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10391976/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9925709","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 1
Role of cellulose response transporter-like protein CRT2 in cellulase induction in Trichoderma reesei. 纤维素反应转运体样蛋白 CRT2 在毛霉菌纤维素酶诱导过程中的作用
Biotechnology for Biofuels and Bioproducts Pub Date : 2023-07-24 DOI: 10.1186/s13068-023-02371-7
Su Yan, Yan Xu, Xiao-Wei Yu
{"title":"Role of cellulose response transporter-like protein CRT2 in cellulase induction in Trichoderma reesei.","authors":"Su Yan, Yan Xu, Xiao-Wei Yu","doi":"10.1186/s13068-023-02371-7","DOIUrl":"10.1186/s13068-023-02371-7","url":null,"abstract":"<p><strong>Background: </strong>Induction of cellulase in cellulolytic fungi Trichoderma reesei is strongly activated by cellulosic carbon sources. The transport of cellulosic inducer and the perception of inducing signal is generally considered as the critical process for cellulase induction, that the inducing signal would be perceived by a sugar transporter/transceptor in T. reesei. Several sugar transporters are coexpressed during the induction stage, but which function they serve and how they work collaboratively are still difficult to elucidate.</p><p><strong>Results: </strong>In this study, we found that the constitutive expression of the cellulose response transporter-like protein CRT2 (previously identified as putative lactose permease TRE77517) improves cellulase induction on a cellulose, cellobiose or lactose medium. Functional studies indicate that the membrane-bound CRT2 is not a transporter of cellobiose, lactose or glucose in a yeast system, and it also does not affect cellobiose and lactose utilization in T. reesei. Further study reveals that CRT2 has a slightly similar function to the cellobiose transporter CRT1 in cellulase induction. Overexpression of CRT2 led to upregulation of CRT1 and the key transcription factor XYR1. Moreover, overexpression of CRT2 could partially compensate for the function loss of CRT1 on cellulase induction.</p><p><strong>Conclusions: </strong>Our study uncovers the novel function of CRT2 in cellulase induction collaborated with CRT1 and XYR1, possibly as a signal transductor. These results deepen the understanding of the influence of sugar transporters in cellulase production.</p>","PeriodicalId":9125,"journal":{"name":"Biotechnology for Biofuels and Bioproducts","volume":"16 1","pages":"118"},"PeriodicalIF":0.0,"publicationDate":"2023-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10364367/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9924176","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Integrated transcriptomic and metabolomic analysis reveals the effects of polyploidization on the lignin content and metabolic pathway in Eucalyptus. 综合转录组学和代谢组学分析揭示了多倍体化对桉树木质素含量和代谢途径的影响。
Biotechnology for Biofuels and Bioproducts Pub Date : 2023-07-21 DOI: 10.1186/s13068-023-02366-4
Tingting Xu, Zhao Liu, Dingju Zhan, Zhenwu Pang, Shuwen Zhang, Chenhe Li, Xiangyang Kang, Jun Yang
{"title":"Integrated transcriptomic and metabolomic analysis reveals the effects of polyploidization on the lignin content and metabolic pathway in Eucalyptus.","authors":"Tingting Xu,&nbsp;Zhao Liu,&nbsp;Dingju Zhan,&nbsp;Zhenwu Pang,&nbsp;Shuwen Zhang,&nbsp;Chenhe Li,&nbsp;Xiangyang Kang,&nbsp;Jun Yang","doi":"10.1186/s13068-023-02366-4","DOIUrl":"https://doi.org/10.1186/s13068-023-02366-4","url":null,"abstract":"<p><strong>Background: </strong>Lignin is a major restriction factor for the industrial production of biomass resources, such as pulp and bioenergy. Eucalyptus is one of the most important sources of pulp and bioenergy. After polyploidization, the lignin content of forest trees is generally reduced, which is considered a beneficial genetic improvement. However, the differences in the lignin content between triploid and diploid Eucalyptus and the underlying regulatory mechanism are still unclear.</p><p><strong>Results: </strong>We conducted a comprehensive analysis at the phenotypic, transcriptional and metabolite levels between Eucalyptus urophylla triploids and diploids to reveal the effects of polyploidization on the lignin content and lignin metabolic pathway. The results showed that the lignin content of Eucalyptus urophylla triploid stems was significantly lower than that of diploids. Lignin-related metabolites were differentially accumulated between triploids and diploids, among which coniferaldehyde, p-coumaryl alcohol, sinapaldehyde and coniferyl alcohol had significant positive correlations with lignin content, indicating that they might be primarily contributing metabolites. Most lignin biosynthetic genes were significantly downregulated, among which 11 genes were significantly positively correlated with the lignin content and above metabolites. Furthermore, we constructed a co-expression network between lignin biosynthetic genes and transcription factors based on weighted gene co-expression network analysis. The network identified some putative orthologues of secondary cell wall (SCW)-related transcription factors, among which MYB52, MYB42, NAC076, and LBD15 were significantly downregulated in Eucalyptus urophylla triploids. In addition, potential important transcription factors, including HSL1, BEE3, HHO3, and NAC046, also had high degrees of connectivity and high edge weights with lignin biosynthetic genes, indicating that they might also be involved in the variation of lignin accumulation between triploid and diploid Eucalyptus urophylla.</p><p><strong>Conclusions: </strong>The results demonstrated that some lignin-related metabolites, lignin biosynthetic genes and transcription factors in Eucalyptus urophylla triploids may be relatively sensitive in response to the polyploidization effect, significantly changing their expression levels, which ultimately correlated with the varied lignin content. The analysis of the underlying formation mechanism could provide beneficial information for the development and utilization of polyploid biomass resources, which will be also valuable for genetic improvement in other bioenergy plants.</p>","PeriodicalId":9125,"journal":{"name":"Biotechnology for Biofuels and Bioproducts","volume":"16 1","pages":"117"},"PeriodicalIF":0.0,"publicationDate":"2023-07-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10360242/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9858284","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Thermotolerance improvement of engineered Saccharomyces cerevisiae ERG5 Delta ERG4 Delta ERG3 Delta, molecular mechanism, and its application in corn ethanol production. 转基因酿酒酵母菌ERG5 Delta ERG4 Delta ERG3 Delta的耐热性改良及其在玉米乙醇生产中的应用
Biotechnology for Biofuels and Bioproducts Pub Date : 2023-04-12 DOI: 10.1186/s13068-023-02312-4
Peizhou Yang, Wenjing Wu, Jianchao Chen, Suwei Jiang, Zhi Zheng, Yanhong Deng, Jiuling Lu, Hu Wang, Yong Zhou, Yuyou Geng, Kanglin Wang
{"title":"Thermotolerance improvement of engineered Saccharomyces cerevisiae ERG5 Delta ERG4 Delta ERG3 Delta, molecular mechanism, and its application in corn ethanol production.","authors":"Peizhou Yang,&nbsp;Wenjing Wu,&nbsp;Jianchao Chen,&nbsp;Suwei Jiang,&nbsp;Zhi Zheng,&nbsp;Yanhong Deng,&nbsp;Jiuling Lu,&nbsp;Hu Wang,&nbsp;Yong Zhou,&nbsp;Yuyou Geng,&nbsp;Kanglin Wang","doi":"10.1186/s13068-023-02312-4","DOIUrl":"https://doi.org/10.1186/s13068-023-02312-4","url":null,"abstract":"<p><strong>Background: </strong>The thermotolerant yeast is beneficial in terms of efficiency improvement of processes and reduction of costs, while Saccharomyces cerevisiae does not efficiently grow and ferment at high-temperature conditions. The sterol composition alteration from ergosterol to fecosterol in the cell membrane of S. cerevisiae affects the thermotolerant capability.</p><p><strong>Results: </strong>In this study, S. cerevisiae ERG5, ERG4, and ERG3 were knocked out using the CRISPR-Cas9 approach to impact the gene expression involved in ergosterol synthesis. The highest thermotolerant strain was S. cerevisiae ERG5ΔERG4ΔERG3Δ, which produced 22.1 g/L ethanol at 37 °C using the initial glucose concentration of 50 g/L with an increase by 9.4% compared with the wild type (20.2 g/L). The ethanol concentration of 9.4 g/L was produced at 42 ℃, which was 2.85-fold of the wild-type strain (3.3 g/L). The molecular mechanism of engineered S. cerevisiae at the RNA level was analyzed using the transcriptomics method. The simultaneous deletion of S. cerevisiae ERG5, ERG4, and ERG3 caused 278 up-regulated genes and 1892 down-regulated genes in comparison with the wild-type strain. KEGG pathway analysis indicated that the up-regulated genes relevant to ergosterol metabolism were ERG1, ERG11, and ERG5, while the down-regulated genes were ERG9 and ERG26. S. cerevisiae ERG5ΔERG4ΔERG3Δ produced 41.6 g/L of ethanol at 37 °C with 107.7 g/L of corn liquefied glucose as carbon source.</p><p><strong>Conclusion: </strong>Simultaneous deletion of ERG5, ERG4, and ERG3 resulted in the thermotolerance improvement of S. cerevisiae ERG5ΔERG4ΔERG3Δ with cell viability improvement by 1.19-fold at 42 °C via modification of steroid metabolic pathway. S. cerevisiae ERG5ΔERG4ΔERG3Δ could effectively produce ethanol at 37 °C using corn liquefied glucose as carbon source. Therefore, S. cerevisiae ERG5ΔERG4ΔERG3Δ had potential in ethanol production at a large scale under supra-optimal temperature.</p>","PeriodicalId":9125,"journal":{"name":"Biotechnology for Biofuels and Bioproducts","volume":"16 1","pages":"66"},"PeriodicalIF":0.0,"publicationDate":"2023-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10091661/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9300494","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 3
Genetic markers and tree properties predicting wood biorefining potential in aspen (Populus tremula) bioenergy feedstock. 预测杨树(山杨)生物能源原料木材生物精炼潜力的遗传标记和树木特性。
Biotechnology for Biofuels and Bioproducts Pub Date : 2023-04-10 DOI: 10.1186/s13068-023-02315-1
Sacha Escamez, Kathryn M Robinson, Mikko Luomaranta, Madhavi Latha Gandla, Niklas Mähler, Zakiya Yassin, Thomas Grahn, Gerhard Scheepers, Lars-Göran Stener, Stefan Jansson, Leif J Jönsson, Nathaniel R Street, Hannele Tuominen
{"title":"Genetic markers and tree properties predicting wood biorefining potential in aspen (Populus tremula) bioenergy feedstock.","authors":"Sacha Escamez, Kathryn M Robinson, Mikko Luomaranta, Madhavi Latha Gandla, Niklas Mähler, Zakiya Yassin, Thomas Grahn, Gerhard Scheepers, Lars-Göran Stener, Stefan Jansson, Leif J Jönsson, Nathaniel R Street, Hannele Tuominen","doi":"10.1186/s13068-023-02315-1","DOIUrl":"10.1186/s13068-023-02315-1","url":null,"abstract":"<p><strong>Background: </strong>Wood represents the majority of the biomass on land and constitutes a renewable source of biofuels and other bioproducts. However, wood is recalcitrant to bioconversion, raising a need for feedstock improvement in production of, for instance, biofuels. We investigated the properties of wood that affect bioconversion, as well as the underlying genetics, to help identify superior tree feedstocks for biorefining.</p><p><strong>Results: </strong>We recorded 65 wood-related and growth traits in a population of 113 natural aspen genotypes from Sweden ( https://doi.org/10.5061/dryad.gtht76hrd ). These traits included three growth and field performance traits, 20 traits for wood chemical composition, 17 traits for wood anatomy and structure, and 25 wood saccharification traits as indicators of bioconversion potential. Glucose release after saccharification with acidic pretreatment correlated positively with tree stem height and diameter and the carbohydrate content of the wood, and negatively with the content of lignin and the hemicellulose sugar units. Most of these traits displayed extensive natural variation within the aspen population and high broad-sense heritability, supporting their potential in genetic improvement of feedstocks towards improved bioconversion. Finally, a genome-wide association study (GWAS) revealed 13 genetic loci for saccharification yield (on a whole-tree-biomass basis), with six of them intersecting with associations for either height or stem diameter of the trees.</p><p><strong>Conclusions: </strong>The simple growth traits of stem height and diameter were identified as good predictors of wood saccharification yield in aspen trees. GWAS elucidated the underlying genetics, revealing putative genetic markers for bioconversion of bioenergy tree feedstocks.</p>","PeriodicalId":9125,"journal":{"name":"Biotechnology for Biofuels and Bioproducts","volume":"16 1","pages":"65"},"PeriodicalIF":0.0,"publicationDate":"2023-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10088276/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9289611","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
An attempt to simultaneously quantify the polysaccharide, total lipid, protein and pigment in single Cyclotella cryptica cell by Raman spectroscopy. 试图用拉曼光谱法同时定量单个隐环菌细胞中的多糖、总脂质、蛋白质和色素。
Biotechnology for Biofuels and Bioproducts Pub Date : 2023-04-08 DOI: 10.1186/s13068-023-02314-2
Xiufen Wang, Yuehui He, Yuanyuan Zhou, Baohua Zhu, Jian Xu, Kehou Pan, Yun Li
{"title":"An attempt to simultaneously quantify the polysaccharide, total lipid, protein and pigment in single Cyclotella cryptica cell by Raman spectroscopy.","authors":"Xiufen Wang,&nbsp;Yuehui He,&nbsp;Yuanyuan Zhou,&nbsp;Baohua Zhu,&nbsp;Jian Xu,&nbsp;Kehou Pan,&nbsp;Yun Li","doi":"10.1186/s13068-023-02314-2","DOIUrl":"https://doi.org/10.1186/s13068-023-02314-2","url":null,"abstract":"<p><strong>Background: </strong>At present, the conventional methods for determining photosynthetic products of microalgae are usually based on a large number of cell mass to reach the measurement baseline, and the result can only reveal the average state at the population level, which is not feasible for large-scale and rapid screening of specific phenotypes from a large number of potential microalgae mutants. In recent years, single-cell Raman spectra (SCRS) has been proved to be able to rapidly and simultaneously quantify the biochemical components of microalgae. However, this method has not been reported to analyze the biochemical components of Cyclotella cryptica (C. cryptica). Thus, SCRS was first attempt to determine these four biochemical components in this diatom.</p><p><strong>Results: </strong>The method based on SCRS was established to simultaneously quantify the contents of polysaccharide, total lipids, protein and Chl-a in C. cryptica, with thirteen Raman bands were found to be the main marker bands for the diatom components. Moreover, Partial Least Square Regression (PLSR) models based on full spectrum can reliably predict these four cellular components, with Pearson correlation coefficient for these components reached 0.949, 0.904, 0.801 and 0.917, respectively. Finally, based on SCRS data of one isogenic sample, the pairwise correlation and dynamic transformation process of these components can be analyzed by Intra-ramanome Correlation Analysis (IRCA), and the results showed silicon starvation could promote the carbon in C. cryptica cells to flow from protein and pigment metabolism to polysaccharide and lipid metabolism.</p><p><strong>Conclusions: </strong>First, method for the simultaneous quantification of the polysaccharide, total lipid, protein and pigment in single C. cryptica cell are established. Second, the instant interconversion of intracellular components was constructed through IRCA, which is based on data set of one isogenic population and more precision and timeliness. Finally, total results indicated that silicon deficiency could promote the carbon in C. cryptica cells to flow from protein and pigment metabolism to polysaccharide and lipid metabolism.</p>","PeriodicalId":9125,"journal":{"name":"Biotechnology for Biofuels and Bioproducts","volume":"16 1","pages":"63"},"PeriodicalIF":0.0,"publicationDate":"2023-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10082982/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9282337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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