Didepside Formation by the Nonreducing Polyketide Synthase Preu6 of Preussia isomera Requires Interaction of Starter Acyl Transferase and Thioesterase Domains

Orsellinic acid (OA) derivatives are produced by filamentous fungi using nonreducing polyketide synthases (nrPKSs). The chain-releasing thioesterase (TE) domains of such nrPKSs were proposed to also catalyze dimerization to yield didepsides, such as lecanoric acid. Here, we use combinatorial domain exchanges, domain dissections and reconstitutions to reveal that the TE domain of the lecanoric acid synthase Preu6 of Preussia isomera must collaborate with the starter acyl transferase (SAT) domain from the same nrPKS. We show that artificial SAT-TE fusion proteins are highly effective catalysts and reprogram the ketide homologation chassis to form didepsides. We also demonstrate that dissected SAT and TE domains of Preu6 physically interact, and SAT and TE domains of OA-synthesizing nrPKSs may co-evolve. Our work highlights an unexpected domain–domain interaction in nrPKSs that must be considered for the combinatorial biosynthesis of unnatural didepsides, depsidones, and diphenyl ethers.     

Polyketide Synthase-Mediated O-Methyloxime Formation in the Biosynthesis of the Oximidine Anticancer Agents

Bacterial trans-acyltransferase polyketide synthases (trans-AT PKSs) are modular megaenzymes that employ unusual catalytic domains to assemble diverse bioactive natural products. One such PKS is responsible for the biosynthesis of the oximidine anticancer agents, oxime-substituted benzolactone enamides that inhibit vacuolar H⁺-ATPases. Here, we describe the identification of the oximidine gene cluster in Pseudomonas baetica and the characterization of four novel oximidine variants, including a structurally simpler intermediate that retains potent anticancer activity. Using a combination of in vivo, in vitro and computational approaches, we experimentally elucidate the oximidine biosynthetic pathway and reveal an unprecedented mechanism for O-methyloxime formation. We show that this process involves a specialized monooxygenase and methyltransferase domain and provide insight into their activity, mechanism and specificity. Our findings expand the catalytic capabilities of trans-AT PKSs and identify potential strategies for the production of novel oximidine analogues.     

The Molecular Basis of Catalysis by SDR Family Members Ketoacyl-ACP Reductase FabG and Enoyl-ACP Reductase FabI in Type-II Fatty Acid Biosynthesis

The short-chain dehydrogenase/reductase (SDR) superfamily members acyl-ACP reductases FabG and FabI are indispensable core enzymatic modules and catalytic orientation controllers in type-II fatty acid biosynthesis. Herein, we report their distinct substrate allosteric recognition and enantioselective reduction mechanisms. FabG achieves allosteric regulation of ACP and NADPH through ACP binding across two adjacent FabG monomers, while FabI follows an irreversible compulsory order of substrate binding in that NADH binding must precede that of ACP on a discrete FabI monomer. Moreover, FabG and FabI utilize a backdoor residue Phe187 or a “rheostat” α8 helix for acyl chain length selection, and their corresponding triad residues Ser142 or Tyr145 recognize the keto- or enoyl-acyl substrates, respectively, facilitating initiation of nucleophilic attack by NAD(P)H. The other two triad residues (Tyr and Lys) mediate subsequent proton transfer and (R)-3-hydroxyacyl- or saturated acyl-ACP production.     

Biosynthesis of Bixa orellana seed extract mediated silver nanoparticles with moderate antioxidant,antibacterial and antiproliferative activity

Biosynthesis of metallic silver nanoparticles (AgNPs) has gained much interest and offers an attractive alternate to physical and chemical approaches. In recent year several safe, easy, cost-effective, reproducible, and environmentally friendly synthesis approaches for silver nanoparticles have been developed. In this research work, a simple, cheap, and unexplored method was applied on green synthesis of AgNPs using secondary metabolites extracted from Bixa orellana seeds. The seeds are rich of flavonoids and phenolic compounds which presumably responsible for the fast reduction and stabilization of silver ion into silver nanoparticles. The biosynthesis process is very likely to be able to reduce silver ions under simple physiological conditions. The surface plasmon resonance (SPR) that was appeared at 420 nm in UV–vis spectrum, had confirmed the formation of AgNPs. Moreover, the functional groups in secondary metabolite that act as reducing, capping and stabilizing agents for silver nanoparticles, are identified by Fourier transform infrared (FTIR) spectra. An X-ray diffraction analysis generated four peaks for Bixa orellana seed extract mediated AgNPs positioned at 2θ angles of 38.1°, 44.2°, 64.6°, and 77.5° corresponding to crystal planes (1 1 1), (2 0 0), (2 2 0), and (3 1 1). Field emission scanning electron microscope (FESEM) and transmission electron microscopy (TEM) images confirmed the formation of nanosized silver particles. The z-average of the synthesized particles measured by dynamic light scattering (DLS) was found to be 92.9 nm. AgNPs synthesized exhibited remarkable antioxidant activity, antibacterial and antiproliferative activity against human breast (MCF-7) cell line. On the basis of our results, we conclude that biologically synthesized AgNPs exhibited favorable characteristics and have the potential to be used in biomedical fields.     

Mevalonate-Independent Biosynthesis of Terpenoid Volatiles in Plants: Induced and Constitutive Emission of Volatiles

Terpenoids from flower and leaf volatiles are predominantly synthesized along the mevalonate-independent pathway, as shown by administration of [D₂]deoxy-D -xylulose and [D₅]mevalolactone. The parallel use of two pathways for the biosynthesis of the sesquiterpenoid-derived 4,8-dimethylnona-1,3,7-triene (DMNT, see picture) may be important to ensure the synthesis of the volatile alarm codes of plants.     

Reverse versus Normal Prenyl Transferases in Paraherquamide Biosynthesis Exhibit Distinct Facial Selectivities

Both a face-selective and a non-face-selective mode of formation of quaternary centers of isoprene-derived structural moieties of the natural alkaloid paraherquamide A ( 1 ) have been discovered by feeding experiments on Penicillium fellutanum with [U-¹³C₆]-glucose and [¹³C₂]-acetate. The labeling patterns suggest that the methyl groups (C22, C23) are introduced in a non-face-selective manner by a reverse prenyl transferase. The C₅ unit comprising the dioxepin moiety retains stereochemical integrity indicative of a single, face-selective addition of the phenolic group to the dimethylallyl group.     

黄钾铁矾的生物合成与鉴定

利用氧化亚铁硫杆菌的生物催化氧化作用,在FeSO4—K2SO4—H2O体系中和常温常压条件下合成赭黄色的黄钾铁矾。借助SEM,XRD,FTIR,ICP—AES等方法对它的化学组成和结构进行了分析与表征。结果表明,Thiobacillus ferrooxidans休止细胞可在2天内将FeSO4-K2SO4-H2O体系中的Fe^2 全部氧化为Fe^3 ,Fe^3 在高浓度硫酸根、K^ 存在和酸性条件下水解生成赭黄色高铁沉淀,经鉴定为黄钾铁矾,其晶体粒径均匀,分散性好,且没有无定形的羟基硫酸高铁副产物。     

Fungal based synthesis of silver nanoparticles—An effect of temperature on the size of particles

A simple and effective approach to aqueous based biosynthesis of silver nanoparticles was demonstrated and the effect of temperature on controlling size of silver nanoparticles was studied. The morphology and uniformity of silver nanoparticles were investigated by UV–Vis spectroscopy, X-ray diffraction and HrTEM. The functional group of protein molecule was identified using FTIR. Increase in reaction temperature leads to decrease in size of silver nanoparticles and increase in monodispersity.     

CdS量子点的酿酒酵母仿生合成及光谱表征

以酿酒酵母为载体,常温下利用仿生法成功合成了CdS量子点。荧光发射光谱、紫外吸收光谱以及荧光显微镜照片证明,该方法合成的CdS量子点的荧光发射峰位置在443nm,在紫外灯下能发蓝绿色荧光。透射电子显微镜(TEM)表征结果表明,该仿生法合成的CdS量子点为六方纤锌矿结构。以荧光发射和紫外吸收光谱为性能指标,考察了酿酒酵母生长时期、Cd2+的反应浓度以及反应时间等条件对合成CdS量子点的影响。当酿酒酵母处于生长稳定期初期时,与浓度为0.5mmol.L-1的Cd2+共培养24h后所合成的CdS量子点荧光最强。实验中观察到,换液培养可有效提高酿酒酵母合成CdS量子点的产量。