Overexpression and repression of key rate-limiting enzymes (acetyl CoA carboxylase and HMG reductase) to enhance fatty acid production from Rhodotorula mucilaginosa.
Acetyl-CoA Carboxylase
/ genetics
Biomass
Carotenoids
/ metabolism
Fatty Acids
/ metabolism
Gene Expression
/ drug effects
Hydroxymethylglutaryl CoA Reductases
/ genetics
Lipid Metabolism
/ drug effects
Metabolic Engineering
Rhodotorula
/ drug effects
Rosuvastatin Calcium
/ pharmacology
Sodium Citrate
/ pharmacology
ACC1
HMG CoA reductase
Rhodotorula mucilaginosa
inducer
inhibitor
Journal
Journal of basic microbiology
ISSN: 1521-4028
Titre abrégé: J Basic Microbiol
Pays: Germany
ID NLM: 8503885
Informations de publication
Date de publication:
Jan 2021
Jan 2021
Historique:
received:
30
06
2020
revised:
05
08
2020
accepted:
27
08
2020
pubmed:
9
9
2020
medline:
30
10
2021
entrez:
8
9
2020
Statut:
ppublish
Résumé
Implementing two-way strategies to enhance the lipid production in Rhodotorula mucilaginosa with the help of metabolic engineering was focused on the overexpression of acetyl coenzyme A carboxylase (ACC1 carboxylase) gene and repression of 3-hydroxy 3-methylglutaryl reductase (HMG-CoA reductase). Using an inducer (sodium citrate) and inhibitor (rosuvastatin), the amounts of biomass, lipid, and carotenoid were estimated. In the presence of inhibitor (200 mM), 62% higher lipid concentration was observed, while 44% enhancement was recorded when inducer (3 mM) was used. A combination of both inhibitor and inducer resulted in a 57% increase in lipid concentration by the oleaginous yeast. These results were again confirmed by real-time polymerase chain reaction by targeting the expression of the genes coding for ACC1 carboxylase and 13-fold increase was recorded in the presence of inducer as compared with control. This combined strategy (inducer and inhibitor use) has been reported for the first time as far as the best of our knowledge. The metabolic engineering strategies reported here will be a powerful approach for the enhanced commercial production of lipids.
Identifiants
pubmed: 32896907
doi: 10.1002/jobm.202000407
doi:
Substances chimiques
Fatty Acids
0
Sodium Citrate
1Q73Q2JULR
Carotenoids
36-88-4
Rosuvastatin Calcium
83MVU38M7Q
Hydroxymethylglutaryl CoA Reductases
EC 1.1.1.-
Acetyl-CoA Carboxylase
EC 6.4.1.2
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
4-14Subventions
Organisme : University Grants Commission
ID : F.15-1/2017/PDFWM-2017-18-DEL-3915(SA-II)
Informations de copyright
© 2020 Wiley-VCH GmbH.
Références
Ward VCA, Munch G, Cicek N, Rehmann L. Direct conversion of the oleaginous yeast Rhodosporidium diobovatum to biodiesel using the ionic liquid [C2mim] [EtSO4]. ACS Sustain. Chem Eng. 2017;5:5562-70.
Chaturvedi S, Bhatacharya A, Nain L, Prasanna R, Khare SK. Valorization of agro-starchy wastes as substrates for oleaginous microbes. Biomass Bioenergy. 2019;127:105294.
Patel A, Mikes F, Matsakas L. An overview of current pretreatment methods used to improve lipid extraction from oleaginous microorganisms. Molecules. 2018;23:1562.
Schirmer A, Rude MA, Li X, Popova E, del Cardayre SB. Microbial biosynthesis of alkanes. Science. 2010;329:559-62.
Zhou YJ, Buijs NA, Zhu Z, Qin J, Siewers V, Nielsen J. Production of fatty acid-derived oleochemicals and biofuels by synthetic yeast cell factories. Nat Commun. 2016;7:11709.
Feng X, Lian J, Zhao H. Metabolic engineering of Saccharomyces cerevisiae to improve 1-hexadecanol production. Metab Eng. 2015;27:10-9.
Nawabi P, Bauer S, Kyrpides N, Lykidis A. Engineering Escherichia coli for biodiesel production utilizing a bacterial fatty acid methyltransferase. Appl Environ Microbiol. 2011;77:8052-61.
Shi S, Valle-Rodriguez JO, Khoomrung S, Siewers V, Nielsen J. Functional expression and characterization of five wax ester synthases in Saccharomyces cerevisiae and their utility for biodiesel production. Biotechnol Biofuels. 2012;5:7-16.
Ma T, Shi B, Ye Z, Li X, Liu M, Chen Y, et al. Lipid engineering combined with systematic metabolic engineering of Saccharomyces cerevisiae for high-yield production of lycopene. Metab Eng. 2019;52:134-42.
Johnravindar D, Karthikeyan OP, Selvam A, Murugesan K. Lipid accumulation potential of oleaginous yeasts: a comparative evaluation using food waste leachate as a substrate. Bioresour Technol. 2018;248:221-8.
Chaturvedi S, Bhattacharya A, Khare SK. Trends in oil production from oleaginous yeast using biomass: biotechnological potential and constraints. Appl Biochem Microbiol. 2018;54:361-9.
Chaturvedi S, Kumari A, Nain L, Khare SK. Bioprospecting microbes for single cell oil production from starchy wastes. Prep Biochem Biotechnol. 2018;48:296-302.
Chaturvedi S, Kumari A, Bhattacharya A, Sharma A, Nain L, Khare SK. Banana peel waste management for single cell oil production. Energy Ecol Environ. 2018;3:296-303.
Thangavelu K, Sundararaju P, Srinivasan N. Simultaneous lipid production for biodiesel feedstock and decontamination of sago processing wastewater using Candida tropicalis ASY2. Biotechnol Biofuels. 2020;13:35.
Tanimura A, Takashima M, Sugita T, Endoh R, Kikukawa M, Yamaguchi S. Cryptococcus terricola is a promising oleaginous yeast for biodiesel production from starch through consolidated bioprocessing. Sci Rep. 2014;4:4776.
Srisuwan W, Techapun C, Seesuriyachan P, Watanabe M, Chaiyaso T. Screening of oleaginous yeast for lipid using rice residue from food waste as a carbon source. KKU Res J. 2016;21:116-26.
Yu A, Zhao Y, Li J. Sustainable production of FAEE biodiesel using the oleaginous yeast Yarrowia lipolytica. Microbiol Open. 2020;9:e1051. https://doi.org/10.1002/mbo3.1051
Dourou M, Aggeli D, Papanikolaou S, Aggelis G. Critical steps in carbon metabolism affecting lipid accumulation and their regulation in oleaginous microorganisms. Appl Microbiol Biotechnol. 2018;102:2509-23.
Wang J, Xu R, Wang R, Haque ME, Liu A. Overexpression of ACC gene from oleaginous yeast Lipomyces starkeyi enhanced the lipid accumulation in Saccharomyces cerevisiae with increased levels of glycerol 3-phosphate substrates. Biosci Biotechnol Biochem. 2016;80:1214-22.
Kim CW, Moona YA, Parka SW, Cheng D, Kwon HJ, Hortona JD. Induced polymerization of mammalian acetyl-CoA carboxylase by MIG12 provides a tertiary level of regulation of fatty acid synthesis. Proc Natl Acad Sci U S A. 2010;107:9626-31.
Sarfaraz RM, Ahmad M, Mahmood A, Minhas MU, Yaqoob A. Development and evaluation of rosuvastatin calcium based microparticles for solubility enhancement: an in vitro study. Adv Polym Technol. 2017;36:433-41.
Galgóczy L, Nyilasi I, Papp T, Vágvölgyi C. Statins as antifungal agents. World J Clin Infect Dis. 2011;30:4-10.
Manzoni M, Rollini M. Biosynthesis and biotechnological production of statins by filamentous fungi and application of these cholesterol-lowering drugs. Appl Microbiol Biotechnol. 2002;58:555-64.
Burg JS, Espenshade PJ. Regulation of HMG-CoA reductase in mammals and yeast. Prog Lipid Res. 2011;50:403-10.
Kurtzman CP, Fell JW, Boekhout T. The yeasts: a taxonomic study. 5th ed. Amsterdam: Elsevier; 2011.
Chaturvedi S, Tiwari R, Bhattacharya A, Nain L, Khare SK. Production of single cell oil by using cassava peel substrate from oleaginous yeast Rhodotorula glutinis. Biocatal Agric Biotechnol. 2019;21:101308.
Gupta VK, Tuohy MG, Ayyachamy M, Turner KM, O'Donovan A. Laboratory protocol in fungal biology. New York: Springer Science and Business Media, Verlag; 2012.
Harrigan WF, McCance ME. Laboratory methods in microbiology. London, UK: Academic Press; 1966. p. 342.
Poli JS, Dallé P, Senter L, Mendes S, Ramirez M, Vainstein MH, et al. Fatty acid methyl ester produced by oleaginous yeast Yarrowia lipolytica QU21: an alternative for vegetable oils. Rev Bras Biosci. 2013;11:203-8.
Kilcawley KN, Wilkinson MG, Fox PF. Determination of key enzyme activities in commercial peptidase and lipase preparations from microbial or animal sources. Enzyme Microb Technol. 2002;31:310-20.
Mokhtari M, Etebarian H, Mirhendi S, Razavi M. Identification and phylogeny of some species of the genera Sporidiobolus and Rhodotorula using analysis of the 5.8s rDNA gene and ribosomal internal transcribed spacers. Arch Biol Sci (Belgrade). 2011;63:79-88.
Sun HY, Singh N. Antimicrobial and immunomodulatory attributes of statins: relevance in solid-organ transplant recipients. Arch Clin Infect Dis. 2009;48:745-55.
Ma Y, Gao Z, Wang Q, Liu Y. Biodiesels from microbial oils: opportunity and challenges. Bioresour Technol. 2019;263:631-41.
Tehlivets O, Scheuringer K, Kohlwein SD. Fatty acid synthesis and elongation in yeast. Biochim Biophys Acta. 2007;1771:255-70.
Wang J, Zhang B, Chen S. Oleaginous yeast Yarrowia lipolytica mutants with a disrupted fatty acyl-CoA synthetase gene accumulate saturated fatty acid. Process Biochem. 2011;46:1436-41.
Ruenwai R, Cheevadhanarak S, Laoteng K. Overexpression of acetyl-CoA carboxylase gene of Mucor rouxii enhanced fatty acid content in Hansenula polymorpha. Mol Biotechnol. 2009;42:327-32.
Meng X, Yang JM, Cao YJ. Increasing fatty acid production in E. coli by simulating the lipid accumulation of oleaginous microorganisms. J Ind Microbiol Biotechnol. 2011;38:919-25.
Davis MS, Solbiati J, Cronan JE. Overproduction of acetyl-CoA carboxylase activity increases the rate of fatty acid biosynthesis in Escherichia coli. J Biol Chem. 2000;275:28593-8.
Lachenmeier DW, Monakhova YB, Kuballa T, Löbell-Behrends S, Maixner S, Kohl-Himmelseher M, et al. NMR evaluation of total statin content and HMG CoA reductase inhibition in red yeast rice (Monascus spp.) food supplements. Chin Med. 2012;7:8.
Westermeyer C, Macreadie IG. Simvastatin reduces ergosterol levels, inhibits growth and causes loss of mtDNA in Candida glabrata. FEMS Yeast Res. 2007;7:436-41.
Liu HW, Zhao X, Wang FJ. The proteome analysis of oleaginous yeast Lipomyces starkeyi. FEMS Yeast Res. 2011;11:42-51.
Knot M, Gupta R, Barve K, Zinjarde S, Govindwar S, Ravikumar A. Fungal production of single cell oil using untreated copra cake and evaluation of its fuel properties for biodiesel. J Microbiol Biotechnol. 2015;25:459-63.