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Biosynthesis of Triacylglycerol Molecules with a Tailored PUFA Profile in Industrial Microalgae

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Molecular Plant 2019年第4期12卷 474-488页

作者： Yi Xin Chen Shen Yiting She Hong Chen Cong Wang Li Wei Kangsup Yoon Danxiang Han Qiang Hu Jian Xu Single-Cell center CAS Key Laboratory of Biofuels and Shandong Laboratory of Energy Genetics Qingdao Institute of BioEnergy and Bioprocess Technology Chinese Academy of Sciences Qingdao Shandong 266101 China center for microalgal biotechnology and biofuels Institute of Hydrobiology Chinese Academy of Sciences Wuhan Hubei 430072 China Core Laboratory Qingdao Institute of BioEnergy and Bioprocess Technology Chinese Academy of Sciences Qingdao Shandong 266101 China Laboratory for Marine Biology and biotechnology Qingdao National Laboratory for Marine Science and Technology Qingdao Shandong 266237 China University of Chinese Academy of Sciences Beijing 100049 China

The composition of polyun saturated fatty acids (PUFAs) in triacylglycerols (TAGs) is key to health ben efits and for oil applications, yet the underlying genetic mechanism remains poorly understood. In this study, by in si Iico, ex vivo, and in vivo profili ng of type-2 diacylglycerol acyltransferases (DGAT2s) in Nannochloropsis oceanica we revealed two novel PUFA-preferring enzymes that discriminate individual PUFA species in TAG assembly, with NoDGAT2J for linoleic acid (LA) and NoDGAT2K for eicosapentaenoic acid (EPA). The LA and EPA composition of TAG molecules is mediated in vivo via the functional partitioning between NoDGAT2J and 2K, both of which are localized in the chloroplast envelope. By modulating transcript abundance of the DGAT2s, an N. oceanica strain bank was created, where proportions of LA and EPA in TAG vary by 18.7-fold (between 0.21% and 3.92% dry weight) and 34.7-fold (between 0.09% and 3.12% dry weight), respective!y. These findings lay the foundation for producing designer TAG molecules with tailored health benefits or for biofuel applications in industrial microalgae and higher-plant crops.

关键词： microalgae polyunsaturated fatty acids diacylglycerol acyltransferase Nannochloropsis oceanica

Conversion of N-doped biochar from carotenoid-extracted Tetraselmis suecica and its application to produce supercapacitors

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Journal of Environmental Sciences 2025年第5期151卷 410-423页

作者： Se Hun Lee Kang Hyun Lee Sang-hwa Lee Soo Kweon Lee Ok Sung Jeon Young Pyo Jeon Dongpyo Hong Young Joon Yoo Sang Yoon Park Hah Young Yoo Advanced Institute of Convergence Technology(AICT) Seoul National UniversitySuwon 16229Republic of Korea Department of biotechnology Sangmyung University20 Hongjimun 2-GilJongno-GuSeoul 03016Republic of Korea Department of Bio-Convergence Engineering Dongyang Mirae University445-8Gyeongin-roGuro-guSeoul 08221Republic of Korea Fermentation team Lotte R&D Center210 Magokjungang-RoGangseo-GuSeoul 07594Republic of Korea School of Electronic Engineering Kyonggi UniversityGyeonggi-do 16227Republic of Korea

Microalgae are one of the promising feedstocks for biorefinery,contributing significantly to net-zero emissions through carbon capture and ***,the disposal of microalgal byproducts from the manufacturing process causes additional environmental pollution,thus,a new application strategy is *** this study,the Tetraselmis suecica byproduct from the carotenoid extraction process was carbonized and converted into *** converted biochar was proved to be nitrogen-doped biochar(NDB),up to 4.69%,with a specific surface area of 206.59m^(2)/g andwas used as an electrode for a *** NDB electrode(NDB-E)in half-cell showed a maximum specific capacitance of 191 F/*** a full-cell test,the NDB-E exhibited a high energy density of 7.396 Wh/kg and a high-power density of 18,100 W/kg,and maintained specific capacity of 95.5%after charge and discharge of 10,000 *** conclusion,our study demonstrated that the carotenoid-extracted microalgal byproducts are a useful resource for the supercapacitor *** approach is the first to convert *** into active materials for supercapacitors.

关键词： Microalgae Tetraselmis suecica Carbonization N-doped biochar Supercapacitor

Unraveling enhanced membrane lipid biosynthesis in Chlamydomonas reinhardtii starchless mutant sta6 by using an electrospray ionization mass spectrometry-based lipidomics method

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Journal of Oceanology and Limnology 2020年第3期38卷 783-794页

作者： ZANG Zhengrong LI Yanhua HU Qiang HAN Danxiang center for microalgal biotechnology and biofuels Institute of HydrobiologyChinese Academy of SciencesWuhan 430072China Key Laboratory for Algal Biology Institute of HydrobiologyChinese Academy of SciencesWuhan 430072China University of Chinese Academy of Sciences Beijing 100049China State Key Laboratory of Freshwater Ecology and biotechnology Institute of HydrobiologyChinese Academy of SciencesWuhan 430072China Laboratory for Marine Biology and biotechnology Qingdao National Laboratory for Marine Science and TechnologyQingdao 266071China Beijing Key Laboratory of Algae Biomass Microalgae Biotechnology CenterSDIC Biotech Investment Co.Ltd.State Development&Investment Corp.Beijing 100142China

The unicellular green alga Chlamydomonas reinhardtii,a well-established model organism,has been widely used in dissecting glycerolipid metabolism in oxygenating photosynthetic *** previous studies,it has been found that shunting carbon precursors from the starch synthesis pathway can lead to a 10-fold increase in TAG content as compared to the wild type,but it is unknown whether inactivation of AGPase may affect membrane lipids *** study aims to investigate global changes in lipid metabolism and homeostasis in the starchless mutant *** *** utilizing an electrospray ionization/mass spectrometry(ESI/MS)-based lipidomics approach,a total of 105 membrane lipid molecules of *** were resolved,including 16 monogalactosyldiacylglycerol(MGDG),16 digalactosyldiacylglycerol(DGDG),11 phosphatidylglycerol(PG),6 sulfoquinovosyldiacylglycerol(SQDG),49 diacylglyceryl-N,N,N-trimethylhomoserine(DGTS),2 phosphatidylethanolamine(PE),and 5 phosphatidylinositol(PI)*** quantitative results indicated that the membrane lipid profiles were similar between the two *** strains grown under both low-and high-light conditions,but the cellular contents of a great number of lipids were altered in sta6 due to the defect in starch *** low-light conditions,sta6 accumulated more PI,MGDG,DGDG but less amounts of DGTS as compared to *** high light,sta6 cells contained higher content membrane lipids than cc-124,except for PG,which is more or less similar in both *** results demonstrate that the cellular membrane lipid homeostasis underwent profound changes in the starchless mutant,and thereby its physiological impact remains to be explored.

关键词： Chlamydomonas reinhardtii chloroplast lipidomes extraplastidic lipidomes electrospray ionization mass spectrometry

Interfacing biosynthetic CdS with engineered Rhodopseudomonas palustris for efficient visible light-driven CO_(2)-CH_(4) conversion

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Frontiers of Chemical Science and Engineering 2024年第10期18卷 1-12页

作者： Yu Zhang Yulei Qian Zhenye Tong Su Yan Xiaoyu Yong Yang-Chun Yong Jun Zhou Bioenergy Research Institute College of Biotechnology and Pharmaceutical EngineeringNanjing Tech UniversityNanjing 211816China biofuels Institute Jiangsu Collaborative Innovation Center of Technology and Material of Water TreatmentSchool of Emergency Management&School of Environment and Safety EngineeringJiangsu UniversityZhenjiang 212013China

Engineered photosynthetic bacterium Rhodo-pseudomonas palustris is excellent at one-step CO_(2) biomethanation and can use near-infrared light sources,overcoming the limitations of conventional photosynthetic *** current study constructed a biohybrid system that deposited CdS nanoparticles on *** biohybrid system broadens the capture of sustainable solar energy,achieving a 155 nmol-mL-biological CH,production under full visible light irradiation,13.4-fold of that by the pure *** transcriptome profiles revealed that gene expression related to photosynthetic electron transfer chain,nitrogenase,nanofilaments,and redox stress defense was ***,we attributed the much-enhanced CO_(2) biomethanation in the biohybrid system to the remarkable increase in the intracellular reducing power and the stronger rigidity of the cells assisted by photoexcited electrons from CdS *** discovery offers insight and a promising strategy forimproving the current CO_(2)-CH_(4) biomanufacturing system.

关键词： CO_(2)methanation Rhodopseudomonas palustris CdS nanoparticles green catalysis

Producing Designer Oils in Industrial Microalgae by Rational Modulation of Co-evolving Type-2 Diacylglycerol Acyltransferases

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Molecular Plant 2017年第12期10卷 1523-1539页

作者： Yi Xin Yandu Lu Yi-Ying Lee Li Wei Jing Jia Qintao Wang Dongmei Wang Fali Bai Hanhua Hu Qiang Hu Jin Liu Yantao Li Jian Xu Single-Cell center CAS Key Laboratory of Biofuels and Shandong Laboratory of Energy Genetics Qingdao Institute of BioEnergy and Bioprocess TechnologyChinese Academy of Sciences Qingdao Shandong 266101 China Institute of Marine and Environmental Technology University of Maryland Center for Environmental Science and University of Maryland Baltimore CountyBaltimore MD 21202 USA Institute for Food and Bioresource Engineering and Department of Energy and Resource Engineering College of Engineering Peking University Beijing 100871China center for microalgal biotechnology and biofuels Institute of Hydrobiology Chinese Academy of Sciences Wuhan Hubei 430072 China Diatom Biology Group Institute of Hydrobiology Chinese Academy of Sciences Wuhan Hubei 430072 China Core Laboratory Qingdao Institute of BioEnergy and Bioprocess Technology Chinese Academy of Sciences Qingdao Shandong 266101 China University of Chinese Academy of Sciences Beijing 100049 China These authors contributed equally to this article.

microalgal oils, depending on their degree of unsaturation, can be utilized as either nutritional supplements or fuels; thus, a feedstock with genetically designed and tunable degree of unsaturation is desirable to maximize process efficiency and product versatility. Systematic profiling of ex vivo （in yeast）, in vitro, and in vivo activities of type-2 diacylglycerol acyltransferases in Nannochloropsis oceanica （NoDGAT2s or NoDGTTs）, via reverse genetics, revealed that NoDGAT2A prefers saturated fatty acids （SFAs）, NoDGAT2D prefers monounsaturated fatty acids （MUFAs）, and NoDGAT2C exhibits the strongest activity toward polyunsaturated fatty acids （PUFAs）. As NoDGAT2A, 2C, and 2D originated from the green alga, red alga, and eukaryotic host ancestral participants of secondary endosymbiosis, respectively, a mecha- nistic model of oleaginousness was unveiled, in which the indigenous and adopted NoDGAT2s formulated functional complementarity and specific transcript abundance ratio that underlie a rigid SFA：MUFA：PUFA hierarchy in triacylglycerol （TAG）. By rationally modulating the ratio of NoDGAT2A＇：2C^D transcripts, a bank of N. oceanica strains optimized for nutritional supplement or fuel production with a wide range of degree of unsaturation were created, in which proportion of SFAs, MUFAs, and PUFAs in TAG varied by 1.3-, 3.7-, and 11.2-fold, respectively. This established a novel strategy to simultaneously improve productivity and quality of oils from industrial microalgae.

关键词： biofuels degree of unsaturation genetic engineering diacylglycerol acyltransferase Nannochloropsis

PHYSIOLOGICAL INHIBITORY EFFECT OF OCS IN ARACHIDONIC ACID-RICH PARIETOCHLORIS INCISA (TREBOUXIOPHYCEAE, CHLOROPHYTA)

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Chinese Journal of Oceanology and Limnology 2002年第3期20卷 248-255页

作者：刘建国张成武 Zvi Cohen Amos Richmond Research and Development center of Marine biotechnology Institute of Oceanol ogy Chinese Academy of Sciences Qingdao 266071 China Division of life Sciences Nanjing Normal University Nanjing 210097 China microalgal biotechnology The Jacob Blaustein Institute for Desert Research Ben-Gurion University of Negev Sede Boker Campus 84993 Israel

Parietochloris incisa is an arachidonic acid rich snow green alga. The main physiological profiles, such as ash free dry weight (AFDW), chlorophyll, carotenoid, protein and total fatty acids (TFA), in this alga exposed to old culture supernatant (OCS) at the decline phase or its crude ethyl acetate extracts (CEAE) were investigated by using tubular photobioreactors of different diameters. Results showed that both OCS and CEAE had strong inhibitory effect on the above physiological parameters. The longer the culture was exposed to OCS and the more CEAE were added into the algal culture, the more the above physiological properties were inhibited. Arachidonic acid (AA), the dominant component of fatty acids in this alga, was also seriously inhibited with respect to total TFA, AFDW of cell mass, or culture volume, due to a probable reduction of enzymes activities catalyzing chain elongation from C18:1ω9 to AA. These results incontestably evidenced that some CEAE dissolving substances existing in OCS, like auto inhibitors, inhibited P. incisa growth through feedback. Hence, any efficient removal of auto inhibitors from algal culture to decrease their bioactivity could be good for maximal production of desired products like AA.

关键词： arachidonic acid Parietochloris incisa growth auto inhibitor

Genome-wide adenine N6-methylation map reveals epigenomic regulation of lipid accumulation in Nannochloropsis

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Plant Communications 2024年第3期5卷 106-122页

作者： Yanhai Gong Qintao Wang Li Wei Wensi Liang Lianhong Wang Nana Lv Xuefeng Du Jiashun Zhang Chen Shen Yi Xin Luyang Sun Jian Xu Single-Cell center CAS Key Laboratory of BiofuelsShandong Key Laboratory of Energy GeneticsQingdao Institute of Bioenergy and Bioprocess TechnologyChinese Academy of SciencesQingdaoChina Shandong Energy Institute QingdaoChina Qingdao New Energy Shandong Laboratory QingdaoChina University of Chinese Academy of Sciences Beijing 100049China Laboratory for Marine Biology and biotechnology Qingdao National Laboratory for Marine Science and TechnologyQingdao 266237China

Epigenetic marks on histones and DNA,such as DNA methylation at N6-adenine(6mA),play crucial roles in gene expression and genome maintenance,but their deposition and function in microalgae remain largely ***,we report a genome-wide 6mA map for the model industrial oleaginous microalga Nannochloropsis oceanica produced by single-molecule real-time *** in 0.1%of adenines,6mA sites are mostly enriched at the AGGYV motif,more abundant in transposons and 30 untranslated re-gions,and associated with active ***,6mA gradually increases in abundance along the direction of gene transcription and shows special positional enrichment near splicing donor and transcrip-tion termination *** expressed genes tend to show greater 6mA abundance in the gene body than do poorly expressed genes,indicating a positive interaction between 6mA and general transcription ***,knockout of the putative 6mA methylase NO08G00280 by genome editing leads to changes in methylation patterns that are correlated with changes in the expression of molybdenum cofactor,sulfate transporter,glycosyl transferase,and lipase genes that underlie reductions in biomass and oil *** contrast,knockout of the candidate demethylase NO06G02500 results in increased 6mA levels and reduced *** the epigenomic players and their roles in biomass productivity and lipid metabolism lays a foundation for epigenetic engineering of industrial microalgae.

关键词： adenine N6-methylation industrial oleaginous microalgae Nannochloropsis oceanica transcriptional regulation epigenomics

Novel kinetic model for the simulation analysis of the butanol productivity of Clostridium acetobutylicum ATCC 824 under different reactor configurations

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Chinese Journal of Chemical Engineering 2018年第4期26卷 812-821页

作者： Hugo I.Velázquez-Sánchez Ricardo Aguilar-López center of Research and Advances Studies of the National Polytechnic Institute (CINVESTAV) Biotechnology and Bioengineering DepartmentAv. National Polytechnic Institute 2508 Col. San Pedro Zacatenco

Acetone–butanol–ethanol(ABE)fermentation process can be exploited for the generation of butanol as biofuel,however it does need to overcome its low volumetric solvent productivity before it can commercially compete with fossil fuel *** this regard,mathematical modelling and simulation analysis are tools that can serve as the base for process engineering development of biological *** this work,a novel phenomenological kinetic model of Clostridium acetobutylicum ATCC 824 was considered as a benchmark system to evaluate the behaviour of an ABE fermentation under different process configurations using both free and immobilized cells:single stage batch operation,fed-batch,single stage Continuous Stirred Tank Reactor(CSTR)and multistage CSTRs with and without biomass *** proposed model achieved a linear correlation index r^2=0.9952 and r^2=0.9710 over experimental data for free and immobilized cells *** predicted maximum butanol concentration and productivity obtained were 13.08 g·L^(-1)and 1.9620 g·L^(-1)·h^(-1)respectively,which represents an increase of 1.01%and 990%versus the currently developed industrial scale process reported currently into the *** results provide a reliable platform for the design and optimization of the ABE fermentation system and showcase the adequate predictive nature of the proposed model.

关键词： biofuels Butanol Clostridium acetobutylicum Mathematical modelling Process configurations

Scalable and automated CRISPR-based strain engineering using droplet microfluidics

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Microsystems & Nanoengineering 2022年第2期8卷 227-236页

作者： Kosuke Iwai Maren Wehrs Megan Garber Jess Sustarich Lauren Washburn Zachary Costello Peter W.Kim David Ando William R.Gaillard Nathan J.Hillson Paul D.Adams Aindrila Mukhopadhyay Hector Garcia Martin Anup K.Singh Technology Division DOE Joint BioEnergy InstituteEmeryvilleCA 94608USA biotechnology and Bioengineering Department Sandia National LaboratoriesLivermoreCA 94550USA biofuels and Bioproducts Division DOE Joint BioEnergy InstituteEmeryvilleCA 94608USA Biological Systems and Engineering Division Lawrence Berkeley National LaboratoryBerkeleyCA 94720USA Molecular Biophysics and Integrated Bioimaging Division Lawrence Berkeley National LaboratoryBerkeleyCA 94720USA Department of Bioengineering University of CaliforniaBerkeleyCA 94720USA BCAM Basque Center for Applied MathematicsBilbao 48009Spain

We present a droplet-based microfluidic system that enables CRISPR-based gene editing and high-throughput screening on a *** microfluidic device contains a 10x10 element array,and each element contains sets of electrodes for two electric field-actuated operations:electrowetting for merging droplets to mix reagents and electroporation for *** device can perform up to 100 genetic modification reactions in parallel,providing a scalable platform for generating the large number of engineered strains required for the combinatorial optimization of genetic pathways and predictable *** demonstrate the system's capabilities through the CRISPR-based engineering of two test cases:(1)disruption of the function of the enzyme galactokinase(galK)in£coli and(2)targeted engineering of the glutamine synthetase gene(glnA)and the blue-pigment synthetase gene(bpsA)to improve indigoidine production in£coli.

关键词： operations generating fluid