Overexpression and repression of key rate‐limiting enzymes (acetyl CoA carboxylase and HMG reductase) to enhance fatty acid production from Rhodotorula mucilaginosa

Abstract

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.

An Innovative Prosopis cineraria Pod Aqueous Waste as Natural Inhibitor for Enhancing Unsaturated Lipids Production in Yeast Cell Using Banana Peel

“…4a). The similar discoloration was observed when rosuvastatin a chemical inhibitor was used for inhibition of HMG CoA reductase enzyme in previous work [5]. It observed that the biomass (banana peel) was better utilized in the chemical inhibited process as compared to the chemical inhibitor.…”

“…In our previous work, it was reported the chemical inhibitor like NaCl and ionic liquid in isolation or combination were shifted the ux towards the accumulation of a higher percentage of SFA in a yeast cell. This SFA may be provided better protection from cell membrane osmosis [5]. In reverse, the natural inhibitor encouraged yeast for the production of UFA, which may be in uenced by the phytomolecules of P. cineraria aqueous extract.…”

“…Oleaginous yeast like Rhodotorula mucilaginosa (Y-1) was found to promising for lipid production in our earlier studies [2,4,5]. Further enhancing the lipid yield, our study is focused on its metabolic pathway for lipid synthesis, and in this regard, the mevalonate pathway is very important.…”

Effectively Improve the Astaxanthin Production by Combined Additives Regulating Different Metabolic Nodes in Phaffia rhodozyma

“…Combinatorial regulation is a beneficial modification strategy that simultaneously targets multiple metabolic nodes and has been successfully applied in other microorganisms ( Li et al, 2012 ; Chaturvedi et al, 2021 ). When metabolic regulators are combined in the fermentation process, multiple nodes of the synthesis process can be simultaneously regulated to increase the regulation efficiency.…”

The Effects of Carbon Source and Growth Temperature on the Fatty Acid Profiles of Thermobifida fusca

“…Several biotin-containing enzyme systems were identifiable via sequence homology, which could generate the malonyl-CoA substrate that is used by FAS to elongate the fatty acid. These enzymes carboxylate acyl-CoA substrates, including acetyl-CoA, and thereby generate malonyl-CoA ( Nikolau et al, 2003 ), which is considered to be the rate-limiting reaction for fatty acid biosynthesis ( Davis et al, 2000 ; Chaturvedi et al, 2021 ). Prior experimental characterization has identified one of these biotin-containing enzymes as being capable of carboxylating acetyl-CoA, propionyl-CoA, or butyryl-CoA ( Shivaiah et al, 2021 ), but whether T. fusca encodes other proteins that can also carboxylate acetyl-CoA will require similar characterizations.…”