Abstract
Raspberry ketone accounts for the characteristic aroma of the raspberry fruit. A bifunctional enzyme with both chalcone synthase (CHS) and benzalacetone synthase (BAS) activity is thought to play a crucial role in the synthesis of p-hydroxybenzalacetone, yet the in vitro enzymatic properties and reaction products of the CHS/BAS recombinant enzyme from raspberry have not been characterized. In this work, a type III polyketide synthase (PKS) gene (RinPKS1) and its corresponding cDNA were isolated from raspberry. Sequence and phylogenetic analyses demonstrated that RinPKS1 is a CHS. However, functional and enzymatic analyses showed that recombinant RinPKS1 is a bifunctional enzyme with both CHS and BAS activity. RinPKS1 showed some interesting characteristics: (1) no traces of bis-noryangonin and 4-coumaroyltriacetic acid lactone could be detected in the enzyme reaction mixture at different pH values; and (2) recombinant RinPKS1 overexpressed in Escherichia coli effectively yielded p-hydroxybenzalacetone as a dominant product at high pH; however, it effectively yielded naringenin as a dominant product at low pH. Furthermore, 4-coumaroyl-CoA and feruloyl-CoA were the only cinnamoyl-CoA derivatives accepted as starter substrates. RinPKS1 did not accept isobutyryl-CoA, isovaleryl-CoA or acetyl-CoA as substrates.
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Abbreviations
- BAS:
-
Benzalacetone synthase
- CHS:
-
Chalcone synthase
- CoA:
-
Coenzyme A
- IPTG:
-
Isopropyl thio-β-d-galactoside
- LC-ESIMS:
-
Liquid chromatography electron spray ionization mass spectrometry
- OKS:
-
Octaketide synthase
- PKS:
-
Polyketide synthase
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (Nos. 31300620 and 31370674) and Key Natural Science Foundation of Beijing Municipality (No. 5111001), Foundation of Beijing Municipal Education Committee (Nos. KM201210020009, KM201310020002, KM201310020015 and KM201410020001), and Comprehensive Reforming Project to promote talents training of BUA (No. BNRC&CC201406).
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Hui-Li Guo and Ya-Dong Yang are contributed equally to this work.
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Guo, HL., Yang, YD., Ma, YD. et al. A bifunctional type III polyketide synthase from raspberry (Rubus idaeus L.) with both chalcone synthase and benzalacetone synthase activity. J. Plant Biochem. Biotechnol. 26, 80–90 (2017). https://doi.org/10.1007/s13562-016-0365-7
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DOI: https://doi.org/10.1007/s13562-016-0365-7