Part:BBa_K4912008
Key enzymes for synthesizing hydroxytyrosol
Bacillus licheniformis is a Gram positive bacterium that exhibits strong tolerance to various environmental stress factors such as high temperature, high salinity, and organic solvents such as acetoin, phenylethanol, and isobutanol. As a biosafety strain, it can synthesize various substances such as polyglutamic acid, prochemic acid, and bacitracin.And due to the metabolic production of 4-hydroxyphenylpyruvate (4-HPP) by Bacillus licheniformis itself, it can be converted into the precursor substance of hydroxytyrosol - tyrosol through the Ehrlich pathway. The Ehrlich pathway is composed of two enzymes, keto decarboxylase and alcohol dehydrogenase, which can generate various branched chain alcohols and aromatic alcohol compounds.
Bacillus licheniformis itself has endogenous alcohol dehydrogenase YugJ, but lacks keto decarboxylase, requiring heterologous introduction.According to reports, overexpression of heterologous Lactococcus lactis derived strains in Bacillus licheniformis α- The keto decarboxylase KivD has successfully constructed a mixed Ehrlich pathway in Bacillus licheniformis. HpaBC is a two component flavin dependent monooxygenase that widely exists in various microorganisms and undergoes hydroxylation reactions on various aromatic compounds, including tyrosol, naringin, p-coumaric acid, caffeic acid, 3,4-dihydroxyphenylpyruvate, ferulic acid, and coniferol. Research has shown that HpaBC derived from Escherichia coli has a strong catalytic ability towards tyrosol and is used for the synthesis of hydroxytyrosol. Therefore, in this study, we selected the gene hpaBC encoding hydroxylase from Escherichia coli.
We used PbacA and P43 promoters to initiate KivD and HpaBC.After testing, the modified strain has been successfully synthesized.
Although we have successfully constructed an engineering strain that can use glucose as a substrate to produce hydroxytyrosol, the yield of hydroxytyrosol has not yet reached the theoretical conversion rate. Therefore, we plan to optimize the conversion of hydroxytyrosol using different promoters
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1143
Illegal PstI site found at 2354
Illegal PstI site found at 3695 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1143
Illegal PstI site found at 2354
Illegal PstI site found at 3695 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1143
Illegal BglII site found at 99
Illegal BglII site found at 919
Illegal BglII site found at 2370 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1143
Illegal PstI site found at 2354
Illegal PstI site found at 3695 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1143
Illegal PstI site found at 2354
Illegal PstI site found at 3695 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 4150
| biology |