Biosynthesis of Nonribosomal Peptides Chitinimides Reveal a Special Type of Thioesterase Domains.

Non-ribosomal peptide synthetases (NRPSs) and their tailored enzymes have diverse biological functions. In this study, we investigated the biosynthesis and function of chitinimides, which belong to the non-ribosomal peptide (NRP) subfamily featuring a pyrrolidine-containing part (X part) connected to the polypeptide chain via an ester bond. A conserved gene cassette, chmHIJK, is responsible for oxyacylation of the pyrrolidine moiety in the X part. The thioesterase (TE) domain of ChmC (ChmC-TE) catalyzes transesterification reactions with a free X part or methanol as a nucleophilic reagent to form different chitinimides. The crucial amino acid residues in the ChmC-TE domains responsible for the specific recognition of the X part were identified, and they were conserved in all the biosynthetic pathways of this NRP subfamily to form a signature motif, YNHNR, suggesting a special type of TE domain in NRPSs. Chitinimides demonstrate the biological function of promoting the swarming ability of the native producer. This study provides deep insights into the biosynthesis of this special NRP subfamily, and shows that the special TE domain could be used to generate diverse NRPs by combinatorial biosynthesis.

Controllable Exfoliation of MOF-Derived Van Der Waals Superstructure into Ultrathin 2D B/N Co-Doped Porous Carbon Nanosheets: A Superior Catalyst for Ambient Ammonia Electrosynthesis.

The electrocatalytic nitrogen reduction reaction (NRR) to synthesize NH3 under ambient conditions is a promising alternative route to the conventional Haber-Bosch process, but it is still a great challenge to develop electrocatalysts' high Faraday efficiency and ammonia yield. Herein, a facile and efficient exfoliation strategy to synthesize ultrathin 2D boron and nitrogen co-doped porous carbon nanosheets (B/NC NS) via a metal-organic framework (MOF)-derived van der Waals superstructure, is reported. The results of experiments and theoretical calculations show that the doping of boron and nitrogen can modulate the electronic structure of the adjacent carbon atoms; which thus, promotes the competitive adsorption of nitrogen and reduces the energy required for ammonia synthesis. The B/NC NS exhibits excellent catalytic performance and stability in electrocatalytic NRR, with a yield rate of 153.4 µg·h-1 ·mg-1 cat and a Faraday efficiency of 33.1%, which is better than most of the reported NRR electrocatalysts. The ammonia yield of B/NC NS can maintain 92.7% of the initial NRR activity after 48 h stability test. The authors' controllable exfoliation strategy using MOF-derived van der Waals superstructure can provide a new insight for the synthesis of other 2D materials.

Unusual Post-Translational Modifications in the Biosynthesis of Lasso Peptides.

Lasso peptides are a subclass of ribosomally synthesized and post-translationally modified peptides (RiPPs) and feature the threaded, lariat knot-like topology. The basic post-translational modifications (PTMs) of lasso peptide contain two steps, including the leader peptide removal of the ribosome-derived linear precursor peptide by an ATP-dependent cysteine protease, and the macrolactam cyclization by an ATP-dependent macrolactam synthetase. Recently, advanced bioinformatic tools combined with genome mining have paved the way to uncover a rapidly growing number of lasso peptides as well as a series of PTMs other than the general class-defining processes. Despite abundant reviews focusing on lasso peptide discoveries, structures, properties, and physiological functionalities, few summaries concerned their unique PTMs. In this review, we summarized all the unique PTMs of lasso peptides uncovered to date, shedding light on the related investigations in the future.

Adsorption in Reversed Order of C2 Hydrocarbons on an Ultramicroporous Fluorinated Metal-Organic Framework.

Metal-organic frameworks have been widely studied in the separation of C2 hydrocarbons, which usually preferentially bind unsaturated hydrocarbons with the order of acetylene (C2 H2 )>ethylene (C2 H4 )>ethane (C2 H6 ). Herein, we report an ultramicroporous fluorinated metal-organic framework Zn-FBA (H2 FBA=4,4'-(hexafluoroisopropylidene)bis(benzoic acid)), shows a reversed adsorption order characteristic for C2 hydrocarbons, that the uptake for C2 hydrocarbons of the framework and the binding affinity between the guest molecule and the framework follows the order C2 H6 >C2 H4 >C2 H2 . Density-functional theory calculations confirm that the completely reversed adsorption order behavior is attributed to the close van der Waals interactions and multiple cooperative C-H⋅⋅⋅F hydrogen bonds between the framework and C2 H6 . Moreover, Zn-FBA exhibits a high selectivity of about 2.9 for C2 H6 over C2 H4 at 298 K and 1 bar. The experimental breakthrough studies show that the high-purity C2 H4 can be obtained from C2 H6 and C2 H4 mixtures in one step.

Performance of 3D printing biomimetic conch shell and pearl shell hybrid design composites under quasi-static three-point bending load.

The failure process of biomimetic hybrid design composite composed of layers of conch shell and pearl shell was studied through quasi-static three-point bending experiments and numerical simulations. The biomimetic conch shell structure with inclined angles serves as the upper layer of the hybrid structure, while the biomimetic pearl shell structure with traditional brick and mud structure serves as the lower layer of the hybrid structure, forming a hybrid design structure. Four inclined angles were designed for the structural units of the conch shell, namely 15°, 30°, 45°, and 60°. Twenty-four specimens (six specimens for each inclined angle) were prepared using 3D printing technology using both soft and hard matrix materials. The influence of different inclined angles on the fracture strength, fracture toughness, and energy absorption of hybrid design structures was experimentally studied. The biomimetic hybrid design composite specimen with a notch is placed between two supporting rollers, and a loading indenter acts at mid-span. All twenty-four specimens were notched with a triangular tip and a rectangular bottom. A loading rate of 1 mm/min is used to avoid the viscoelastic effect of the composite materials. Details of the specimens, the experimental set-up and procedure are discussed in this paper. Complementary to the experimental studies, an extensive numerical investigation was carried out to study the influence of the aspect ratio of brick and mud units on the fracture initiation and failure of hybrid design structures. The causes of crack initiation and propagation, and failure modes in biomimetic hybrid design structures were postulated. These numerical findings help in reinforcing the experimental results and provide crucial information to enhance further research in this exciting area.

[Risk factors for the kidney stones: a hospital-based case-control study in a distric hospital in Beijing].

OBJECTIVE: To explore the risk and protective factors of kidney calculi in order to put forward theoretical basis for preventive and control measures. METHODS: A 1:1 matched case-control study was performed using data from a hospital in Beijing. The case group included 100 inpatients who were diagnosed kidney calculi using B ultrasonic, X-ray and intravenous pyelography during the survey while other 100 urolithiasis and endocrine disease excluded inpatients who shared the same sex, within five years gap to the case group inpatients were for the control group. A face-to-face survey was conducted with self-made questionnaires which covered demographic characteristics, water issues, dietary habits, genetic and medical history. Epidata 3.0 was used to build the database and SPSS 19.0 for the statistical analysis. RESULTS: In the univariate Logistic regression analysis, ten variables were found showing statistical significance. For the multivariate Logistic regression analysis, variables left in the model were labor intensity (OR=0.622, 95%CI: 0.435-0.889), preferring to drink after dinner (OR=0.316, 95%CI: 0.122-0.815), loving drinking (OR=0.232, 95%CI: 0.084-0.642), drinking tea regularly (OR=1.463, 95%CI: 1.033-2.071), eating more vegetables (OR=0.571, 95%CI: 0.328-0.993), the history of the urolithiasis (OR=2.127, 95%CI: 1.065-90.145). CONCLUSION: Drinking tea regularly, urolithiasis history and brain work are the risk factors of kidney calculi while loving drinking and eating more vegetables for the protection.

Leader peptide removal in lasso peptide biosynthesis based on penultimate isoleucine residue.

Lasso peptides are ribosomally synthesized peptides that undergo post-translational modifications including leader peptide removal by B (or the segregated B1 and B2) proteins and core peptide macrolactamization by C proteins to form a unique lariat topology. A conserved threonine residue at the penultimate position of leader peptide is hitherto found in lasso peptide precursors and shown to be a critical recognition element for effective enzymatic processing. We identified a lasso peptide biosynthetic gene cluster (bsf) from Bradymonas sediminis FA350, a Gram-negative and facultatively prey-dependent bacterium that belongs to a novel bacterial order Bradymonadales in the class Deltaproteobacteria. The kinase BsfK specifically catalyzes the phosphorylation of the precursor peptide BsfA on the Ser3 residue. BsfB1 performs dual functions to accelerate the post-translational phosphorylation and assist BsfB2 in leader peptide removal. Most importantly, the penultimate residue of leader peptide is an isoleucine rather than the conserved threonine and this isoleucine has a marked impact on the phosphorylation of Ser3 as well as leader peptide removal, implying that BsfB1 and BsfB2 exhibit a new substrate selectivity for leader peptide binding and excision. This is the first experimentally validated penultimate isoleucine residue in a lasso peptide precursor to our knowledge. In silico analysis reveals that the leader peptide Ile/Val(-2) residue is rare but not uncommon in phosphorylated lasso peptides, as this residue is also discovered in Acidobacteriaceae and Sphingomonadales in addition to Bradymonadales.

Rational construction of genome-reduced Burkholderiales chassis facilitates efficient heterologous production of natural products from proteobacteria.

Heterologous expression of biosynthetic gene clusters (BGCs) avails yield improvements and mining of natural products, but it is limited by lacking of more efficient Gram-negative chassis. The proteobacterium Schlegelella brevitalea DSM 7029 exhibits potential for heterologous BGC expression, but its cells undergo early autolysis, hindering further applications. Herein, we rationally construct DC and DT series genome-reduced S. brevitalea mutants by sequential deletions of endogenous BGCs and the nonessential genomic regions, respectively. The DC5 to DC7 mutants affect growth, while the DT series mutants show improved growth characteristics with alleviated cell autolysis. The yield improvements of six proteobacterial natural products and successful identification of chitinimides from Chitinimonas koreensis via heterologous expression in DT mutants demonstrate their superiority to wild-type DSM 7029 and two commonly used Gram-negative chassis Escherichia coli and Pseudomonas putida. Our study expands the panel of Gram-negative chassis and facilitates the discovery of natural products by heterologous expression.

Obesity-induced overexpression of miRNA-24 regulates cholesterol uptake and lipid metabolism by targeting SR-B1.

Scavenger Receptor B1 (SR-B1) is an 82 kDa integral membrane glycoprotein that mediates selective uptake of high-density lipoprotein cholesteryl ester (CE) in vitro and in vivo. Previously, we defined several kinds of regulatory mechanisms of SR-B1 expression and function. Here, we have dissected the function of a novel miR-24 on SR-B1 expression, HDL uptake and lipid metabolism. We showed that miR-24 was upregulated in HepG2 cells cultured in the mimicked hyperlipidemic condition and in the livers of dietary induced and genetic obesity mice. Overexpression of miR-24 inhibited SR-B1 expression by directly targeting SR-B1 3' UTR and repressed HDL uptake and steroidogenesis in steroidogenic cells. HepG2 cells with miR-24 showed attenuation of TG levels and lipid accumulation. Moreover, we validated that overexpression of miR-24 downregulated the expression of certain genes involved in lipogenesis, FASN, ACLY and SCD1, and increased the expression of genes of cholesterol synthesis, HMGCR, DHCR24 and SREBP2. Taken together, we demonstrated that obesity induced miR-24 repressed HDL uptake, steroid hormone synthesis and lipid metabolism by targeting SR-B1.