Type II thioesterase from Streptomyces coelicolor A3(2)
               The GenBank accession number for the sequence reported in this paper is AF109727.

Kotowska, Magdalena; Pawlik, Krzysztof; Butler, Andrew R.; Cundliffe, Eric; Takano, Eriko; Kuczek, Katarzyna

doi:10.1099/00221287-148-6-1777

Cited by 28 publications

(27 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The recombinant protein hydrolyzed all substrates tested at significant rates. This is consistent with its editing function suggested by a complementation study of S. fradiae (25). The enzyme showed a clear preference for propionate.…”

Section: Discussionsupporting

confidence: 76%

“…To prove this hypothesis, it is necessary to solve the chemical structure of the polyketide produced by the Cpk synthase and to High activity of ScoT for propionate can explain the results of our previous experiments. When the scoT gene from S. coelicolor A3(2) was used to complement a tylO disruption mutant of S. fradiae, polyketide production was restored to a level that was up to 50% of the level of the wild-type strain (25), while complementation with the native thioesterase gene (tylO) restored polyketide production to a level that was nearly 100% of the level of the wild-type strain (6). The starter unit for tylosin is propionate.…”

Section: Discussionmentioning

confidence: 99%

“…This paper deals with the TE II ScoT associated with the type I modular PKS Cpk from Streptomyces coelicolor A3(2) (32). ScoT was shown previously to restore polyketide production in a mutant of Streptomyces fradiae lacking the native TE II (encoded by tylO in the tylosin synthase cluster) (25). It was not possible to study the role of this enzyme in the natural context, since the polyketide produced by the Cpk synthase has not been identified yet.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Type II Thioesterase ScoT, Associated with Streptomyces coelicolor A3(2) Modular Polyketide Synthase Cpk, Hydrolyzes Acyl Residues and Has a Preference for Propionate

Kotowska

Pawlik

Smulczyk-Krawczyszyn

et al. 2009

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

Type II thioesterases (TE IIs) were shown to maintain the efficiency of polyketide synthases (PKSs) by removing acyl residues blocking extension modules. However, the substrate specificity and kinetic parameters of these enzymes differ, which may have significant consequences when they are included in engineered hybrid systems for the production of novel compounds. Here we show that thioesterase ScoT associated with polyketide synthase Cpk from Streptomyces coelicolor A3(2) is able to hydrolyze acetyl, propionyl, and butyryl residues, which is consistent with its editing function. This enzyme clearly prefers propionate, in contrast to the TE IIs tested previously, and this indicates that it may have a role in control of the starter unit. We also determined activities of ScoT mutants and concluded that this enzyme is an ␣/␤ hydrolase with Ser90 and His224 in its active site.

show abstract

Section: Discussionsupporting

confidence: 76%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Type II Thioesterase ScoT, Associated with Streptomyces coelicolor A3(2) Modular Polyketide Synthase Cpk, Hydrolyzes Acyl Residues and Has a Preference for Propionate

Kotowska

Pawlik

Smulczyk-Krawczyszyn

et al. 2009

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

show abstract

“…Removal of the TEII from the rifamycin assembly line resulted in a 60% decrease in product yield (25). Neither TEIs nor TEIIs may rescue the disrupted function of the other (6), but a TEII from another pathway may rescue the function of a disrupted TEII (26).…”

mentioning

confidence: 99%

Structure and Functional Analysis of RifR, the Type II Thioesterase from the Rifamycin Biosynthetic Pathway

Claxton

Akey

Silver

et al. 2009

Journal of Biological Chemistry

120

View full text Add to dashboard Cite

Two thioesterases are commonly found in natural product biosynthetic clusters, a type I thioesterase that is responsible for removing the final product from the biosynthetic complex and a type II thioesterase that is believed to perform housekeeping functions such as removing aberrant units from carrier domains. We present the crystal structure and the kinetic analysis of RifR, a type II thioesterase from the hybrid nonribosomal peptide synthetases/polyketide synthase rifamycin biosynthetic cluster of Amycolatopsis mediterranei. Steady-state kinetics show that RifR has a preference for the hydrolysis of acyl units from the phosphopantetheinyl arm of the acyl carrier domain over the hydrolysis of acyl units from the phosphopantetheinyl arm of acyl-CoAs as well as a modest preference for the decarboxylated substrate mimics acetyl-CoA and propionyl-CoA over malonyl-CoA and methylmalonyl-CoA. Multiple RifR conformations and structural similarities to other thioesterases suggest that movement of a helical lid controls access of substrates to the active site of RifR.

show abstract

“…However, there might well be a (yet-unknown) transferase which directly transfers long-chain acyl moieties from ACP to CoA. Streptomyces TEs have not been characterized in the context of storage lipid synthesis yet, but the involvement of several TEs in PKS was studied (67)(68)(69). Type II TEs are dislocated from the multimodular PKS machinery and remove acyl residues from the extension modules.…”

Section: Resultsmentioning

confidence: 99%

Role of Wax Ester Synthase/Acyl Coenzyme A:Diacylglycerol Acyltransferase in Oleaginous Streptomyces sp. Strain G25

Röttig

Strittmatter

Schauer

et al. 2016

Appl Environ Microbiol

View full text Add to dashboard Cite

Recently, we isolated a novel Streptomyces strain which can accumulate extraordinarily large amounts of triacylglycerol (TAG) and consists of 64% fatty acids (dry weight) when cultivated with glucose and 50% fatty acids (dry weight) when cultivated with cellobiose. To identify putative gene products responsible for lipid storage and cellobiose utilization, we analyzed its draft genome sequence. A single gene encoding a wax ester synthase/acyl coenzyme A (CoA):diacylglycerol acyltransferase (WS/DGAT) was identified and heterologously expressed in Escherichia coli. The purified enzyme Atf G25 showed acyltransferase activity with C 12 -or C 16 -acyl-CoA, C 12 to C 18 alcohols, or dipalmitoyl glycerol. This acyltransferase exhibits 24% amino acid identity to the model enzyme AtfA from Acinetobacter baylyi but has high sequence similarities to WS/DGATs from other Streptomyces species. To investigate the impact of Atf G25 on lipid accumulation, the respective gene, atf G25 , was inactivated in Streptomyces sp. strain G25. However, cells of the insertion mutant still exhibited DGAT activity and were able to store TAG, albeit in lower quantities and at lower rates than the wild-type strain. These findings clearly indicate that Atf G25 has an important, but not exclusive, role in TAG biosynthesis in the novel Streptomyces isolate and suggest the presence of alternative metabolic pathways for lipid accumulation which are discussed in the present study. IMPORTANCEA novel Streptomyces strain was isolated from desert soil, which represents an extreme environment with high temperatures, frequent drought, and nutrient scarcity. We believe that these harsh conditions promoted the development of the capacity for this strain to accumulate extraordinarily large amounts of lipids. In this study, we present the analysis of its draft genome sequence with a special focus on enzymes potentially involved in its lipid storage. Furthermore, the activity and importance of the detected acyltransferase were studied. As discussed in this paper, and in contrast to many other bacteria, streptomycetes seem to possess a complex metabolic network to synthesize lipids, whereof crucial steps are still largely unknown. This paper therefore provides insights into a range of topics, including extremophile bacteria, the physiology of lipid accumulation, and the biotechnological production of bacterial lipids. Even extreme environments, such as arid, desert-like regions, are populated by prokaryotes. These mostly Gram-positive bacteria are subjected to extreme fluctuations of temperature and water supply. In addition to desiccation and the resulting cellular damage, the bacteria frequently encounter nutrient limitation. To withstand these harsh environmental conditions, the majority of bacteria have evolved various strategies and are able to store lipophilic compounds such as polyhydroxyalkanoates (PHA), triacylglycerols (TAG), or wax esters (WE) (1-3). TAG are synthesized as primary storage compounds by many actinomycetes-which represent the pre...

show abstract

Type II thioesterase from Streptomyces coelicolor A3(2) The GenBank accession number for the sequence reported in this paper is AF109727.

Cited by 28 publications

References 30 publications

Type II Thioesterase ScoT, Associated with Streptomyces coelicolor A3(2) Modular Polyketide Synthase Cpk, Hydrolyzes Acyl Residues and Has a Preference for Propionate

Type II Thioesterase ScoT, Associated with Streptomyces coelicolor A3(2) Modular Polyketide Synthase Cpk, Hydrolyzes Acyl Residues and Has a Preference for Propionate

Structure and Functional Analysis of RifR, the Type II Thioesterase from the Rifamycin Biosynthetic Pathway

Role of Wax Ester Synthase/Acyl Coenzyme A:Diacylglycerol Acyltransferase in Oleaginous Streptomyces sp. Strain G25

Contact Info

Product

Resources

About