Bacterial Product Template Domain: An Evolutionary Intermediate between Dehydratase and Aldol Cyclase of Type I Polyketide Synthases

The product template (PT) domains act as an aldol cyclase to control the regiospecific aldol cyclization of the extremely reactive poly-β-ketone intermediate assembled by an iterative type I polyketide synthases (PKSs). Up to now, only the structure of fungal PksA PT that mediates the first-ring cyclization via C4-C9 aldol cyclization is available. We describe here the structural and computational characterization of a bacteria PT domain that controls C2-C7 cyclization in orsellinic acid (OSA) synthesis. Mutating the catalytic His949 of the PT abolishes production of OSA and results in a tetraacetic acid lactone (TTL) generated by spontaneous O-C cyclization of the acyl carrier protein (ACP)-bound tetraketide intermediate. Crystal structure of the bacterial PT domain closely resembles dehydrase (DH) domains of modular type I PKSs in the overall fold, dimerization interface and catalytic “His-Asp” dyad organization, but is significantly different from PTs of fungal iterative type I PKSs. QM/MM calculation reveals that the catalytic His949 abstracts a proton from C2 and transfers it to C7 carbonyl to mediate the cyclization reaction. According to the structural similarity to DHs and the functional similarity to fungal PTs, we propose that the bacterial PT represents an evolutionary intermediate between the two tailoring domains of type I PKSs.

Preparation and Performance Evaluation of Gemini Sulphobetaine Surfactant Bis{[(N-methyl-N-(3-alkoxy-2-hydroxy) propyl-N-(3-sulfonic) propyl] methylene}

Surfactants are very often used in tertiary oil production. However, the conventional surfactants cannot be used in high salinity reservoirs of tertiary oil production because the high salinity leads to the deactivation of the surfactants. Gemini sulpho-betaine surfactants have been developed to meet the requirements of tertiary oil production in such reservoirs. In this study, the Gemini sulphobetaine surfactant (bis{[(N-methyl-N-(3-alkoxy-2-hydroxy)propyl-N-(3-sulfonate)propyl]methylene}) was prepared by etherification, ring opening reaction and sulfonation reaction with decyl alcohol, epichlorohydrin, N,N’-dimethyl ethylene diamine and 1,3-propanesultone. The experimental conditions obtained are: the ratio of 1,3-propanesulfonic acid lactone to tertiary amine intermediate = 2.3 : 1, reaction temperature = 70°C and reaction time = 11 h. The analysis of the infrared spectrum showed that the structure of the synthesised substance corresponds to that of a Gemini sulphobetaine surfactant. The chemical shifts of the groups were determined by 1HNMR structural characterisation of the products. The surface tension, emulsifying properties, foaming properties and wetting properties were investigated. The results showed that the surfactant has good foaming properties and good compatibility in a high salt environment.

Sorting for secreted molecule production using a biosensor-in-microdroplet approach

Sorting large libraries of cells for improved small molecule secretion is throughput limited. Here, we combine producer/secretor cell libraries with whole-cell biosensors using a microfluidic-based screening workflow. This approach enables a mix-and-match capability using off-the-shelf biosensors through either coencapsulation or pico-injection. We demonstrate the cell type and library agnostic nature of this workflow by utilizing single-guide RNA, transposon, and ethyl-methyl sulfonate mutagenesis libraries across three distinct microbes (Escherichia coli, Saccharomyces cerevisiae, and Yarrowia lipolytica), biosensors from two organisms (E. coli and S. cerevisiae), and three products (triacetic acid lactone, naringenin, and L-DOPA) to identify targets improving production/secretion.

Complete and efficient conversion of plant cell wall hemicellulose into high-value bioproducts by engineered yeast

Plant cell wall hydrolysates contain not only sugars but also substantial amounts of acetate, a fermentation inhibitor that hinders bioconversion of lignocellulose. Despite the toxic and non-consumable nature of acetate during glucose metabolism, we demonstrate that acetate can be rapidly co-consumed with xylose by engineered Saccharomyces cerevisiae. The co-consumption leads to a metabolic re-configuration that boosts the synthesis of acetyl-CoA derived bioproducts, including triacetic acid lactone (TAL) and vitamin A, in engineered strains. Notably, by co-feeding xylose and acetate, an enginered strain produces 23.91 g/L TAL with a productivity of 0.29 g/L/h in bioreactor fermentation. This strain also completely converts a hemicellulose hydrolysate of switchgrass into 3.55 g/L TAL. These findings establish a versatile strategy that not only transforms an inhibitor into a valuable substrate but also expands the capacity of acetyl-CoA supply in S. cerevisiae for efficient bioconversion of cellulosic biomass.

Structure modification, antialgal, antiplasmodial, and toxic evaluations of a series of new marine-derived 14-membered resorcylic acid lactone derivatives

Marine natural products play critical roles in the chemical defense of many marine organisms and are essential, reputable sources of successful drug leads. Sixty-seven 14-membered resorcylic acid lactone derivatives 3–27 and 30–71 of the natural product zeaenol (1) isolated from the marine-derived fungus Cochliobolus lunatus were semisynthesized by chlorination, acylation, esterification, and acetalization in one to three steps. The structures of these new derivatives were established by HRESIMS and NMR techniques. All the compounds (1–71) were evaluated for their antialgal and antiplasmodial activities. Among them, 14 compounds displayed antifouling activities against adhesion of the fouling diatoms. In particular, 9 and 34 exhibited strong and selective inhibitory effects against the diatoms Navicula laevissima and Navicula exigua (EC50 = 6.67 and 8.55 μmol/L), respectively, which were similar in efficacy to those of the positive control SeaNine 211 (EC50 = 2.90 and 9.74 μmol/L). More importantly, 38, 39, and 69–71 showed potent antiplasmodial activities against Plasmodium falciparum with IC50 values ranging from 3.54 to 9.72 μmol/L. Very interestingly, the five antiplasmodial derivatives displayed non-toxicity in the cytotoxicity assays and the zebrafish embryos model, thus, representing potential promising antiplasmodial drug agents. The preliminary structure–activity relationships indicated that biphenyl substituent at C-2, acetonide at positions C-5′ and C-6′, and tri- or tetra-substituted of acyl groups increased the antiplasmodial activity. Therefore, combining evaluation of chemical ecology with pharmacological models will be implemented as a systematic strategy, not only for environmentally friendly antifoulants but also for structurally novel drugs.

In vitro study for antifungal compounds from Parinari curatellifolia (Chrysobalanaceae) and Terminalia sericea (Combretaceae)

Parinari curatellifolia (Chrysobalanaceae) and Terminalia sericea (Combretaceae) have been traditionally used in Southern Highlands of Tanzania for treatment of various infectious disorders. The present study aimed to evaluate antifungal activity of the isolated compounds from Parinari curatellifolia and Terminalia sericea plant species. The ethyl acetate extract of the root barks from Parinari curatellifolia and Terminalia sericea were fractionated using column chromatography. The structures of compounds were established using both 1D and 2D-NMR spectroscopic techniques while antifungal activities of the fractions and isolated compounds were evaluated using broth microdilution assay against Candida albicans, Cryptococcus neoformans and Aspergillus niger species. Two known compounds toddalolactone (1) and 10-hydroxy-13-methoxy-9- methyl-15-oxo-20-norkaur-16-en-18-oic acid -lactone (2) from P. curatellifolia and two compounds Sericic acid (3) and sericoside (4) from T. sericea were isolated and their structures identified and confirmed by spectral data obtained and from the literatures. Strong antifungal activity was shown by Sericic acid (3) with MIC value of0.07 mg/ml against C. albicans and C. neoformans. Isolation of toddalolactone (1) from Parinari curatellifolia as well as the antifungal activity of Sericic acid (3) from Terminalia sericea is being reported for the first time. Bioactivity of these compounds support traditional use of the studied plants. Keywords: Sericic acid, toddalolactone, fungi, antifungal, Parinari curatellifolia, Terminalia sericea.

Ursolic Acid Lactone Obtained from Eucalyptus tereticornis Increases Glucose Uptake and Reduces Inflammatory Activity and Intracellular Neutral Fat: An In Vitro Study

Obesity has a strong relationship to insulin resistance and diabetes mellitus, a chronic metabolic disease that alters many physiological functions. Naturally derived drugs have aroused great interest in treating obesity, and triterpenoids are natural compounds with multiple biological activities and antidiabetic mechanisms. Here, we evaluated the bioactivity of ursolic acid lactone (UAL), a lesser-known triterpenoid, obtained from Eucalyptus tereticornis. We used different cell lines to show for the first time that this molecule exhibits anti-inflammatory properties in a macrophage model, increases glucose uptake in insulin-resistant muscle cells, and reduces triglyceride content in hepatocytes and adipocytes. In 3T3-L1 adipocytes, UAL inhibited the expression of genes involved in adipogenesis and lipogenesis, enhanced the expression of genes involved in fat oxidation, and increased AMP-activated protein kinase phosphorylation. The range of biological activities demonstrated in vitro indicates that UAL is a promising molecule for fighting diabetes.