Site-saturation mutagenesis is a powerful tool for protein optimization due to its efficiency and simplicity. A degenerate codon NNN or NNS (K) often used encode the 20 standard amino acids, but this will produce redundant codons cause uneven distribution of acids in constructed library. Here we present novel "small-intelligent" strategy construct libraries that have minimal gene library size without inherent acid biases, stop codons, rare Escherichia coli by coupling well-designed...

Thermophilic proteins have various practical applications in theoretical research and industry. In recent years, the demand for thermophilic on an industrial scale has been increasing; therefore, engineering of become a hot direction field protein engineering. However, exact mechanism thermostability is not yet known, knowing basis necessary. order to understand proteins, we made statistical analysis sequences, secondary structures, hydrogen bonds, salt bridges, DHA (Donor–Hydrogen–Accepter)...

Eradicating poverty and improving human well-being are pivotal objectives for achieving global sustainable development. Sustainable agriculture, as a key domain, plays crucial role in addressing poverty. However, to date, there remains insufficient research on the specific impact of agricultural green development To bridge this gap, we utilize panel data from 273 prefecture-level cities China 2006 2022 explore how affects based constructing regional multidimensional index an agriculture. Our...

ABSTRACT Halohydrin dehalogenases, also known as haloalcohol dehalogenases or halohydrin hydrogen-halide lyases, catalyze the nucleophilic displacement of a halogen by vicinal hydroxyl function in halohydrins to yield epoxides. Three novel bacterial genes encoding were cloned and expressed Escherichia coli , enzymes shown display remarkable differences substrate specificity. The dehalogenase Agrobacterium radiobacter strain AD1, designated HheC, was purified homogeneity. k cat K m values...

Agricultural Technology Demonstration Centre (ATDC) has been considered as an alternative model to pursue sustainability of Chinese foreign aid African countries in the new era. This paper attempts examine ATDC scheme, particularly focus on knowledge construction at macro level design and encounter micro daily operation based case studies Tanzania, Ethiopia, Zimbabwe, Mozambique. Our study finds multiple facets ATDC, i.e., cleavage between macro-level policy implementation realities;...

Thermophilic proteins have important application value in biotechnology and industrial processes. The correct identification of thermophilic provides information for the these engineering. method based on biochemistry is laborious, time-consuming, high cost. Therefore, there an urgent need a fast accurate to identify proteins. Considering this urgency, we constructed reliable benchmark dataset containing 1,368 1,443 non-thermophilic A multi-layer perceptron (MLP) model multi-feature fusion...

[figure: see text] The halohydrin dehalogenase from Agrobacterium radiobacter AD1 catalyzed the highly enantioselective and beta-regioselective azidolysis of (substituted) styrene oxides. By means kinetic resolutions remaining epoxide formed azido alcohol could be obtained in very high ee. In a large scale conversion, decrease yield selectivity due to uncatalyzed chemical side reaction overcome by slow addition azide.

Halohydrin dehalogenase from Agrobacterium radiobacter catalyzed the enantioselective ring opening of terminal epoxides with cyanate as a nucleophile, yielding 5-substituted oxazolidinones in high yields and enantiopurity (69−98% ee). This is first example biocatalytic conversion range to corresponding oxazolidinones.

Mozambique, a country undergoing rapid transformations driven by the recent discovery of mineral resources, is one top destinations for Chinese and Brazilian cooperation investment in Africa.This article provides an account policies, narratives, operational modalities underlying motivations development Mozambique.It particularly interested understanding how engagements are perceived talked about, what drives them formal informal relations emerging at level particular exchanges.The draws on...

Halo alcohol dehalogenase HheC catalyzes the highly enantioselective dehalogenation of vicinal halo alcohols to epoxides, as well reverse reaction, and β-regioselective nucleophilic ring opening epoxides by pseudo-halides such azide cyanide. To investigate this latter we determined X-ray structures complexes with favored unfavored enantiomers para-nitrostyrene oxide. The aromatic parts two bind in a very similar way, but epoxide (S)-enantiomer binds nonproductive inverted manner, oxygen Cβ...

Abstract The halohydrin dehalogenase from Agrobacterium radiobacter AD1 (HheC) catalyzes nucleophilic ring opening of epoxides with cyanide and azide. In the case 2,2‐disubstituted epoxides, this reaction proceeds excellent enantioselectivity (E values up to>200), which gives, by kinetic resolution, access to various enantiopure β‐substituted tertiary alcohols ( ee 99 %). Since enzyme has a broad substrate range because these are difficult prepare in other ways, HheC is an attractive...

A novel activity of halohydrin dehalogenases towards spiroepoxides has been found. The enzyme from Arthrobacter sp. (HheA) catalysed highly regioselective azidolysis containing 5, 6 and 7-membered cycloalkane rings, while the Agrobacterium radiobacter (HheC), besides high regioselectivity, also displayed moderate to enantioselectivity (E up >200) that can be applied for kinetic resolution chiral spiroepoxides. orientations in active site were studied by quantum-chemical calculations docking...

ABSTRACT Halohydrin dehalogenase from Agrobacterium radiobacter AD1 (HheC) is a valuable tool in the preparation of R enantiomers epoxides and β-substituted alcohols. In contrast, halohydrin Arthrobacter sp. AD2 (HheA) shows low S enantioselectivity toward most aromatic substrates. Here, three amino acids (V136, L141, N178) located two neighboring active-site loops HheA were proposed to be key residues for controlling enantioselectivity. They subjected saturation mutagenesis aimed at...

Bacteriophages, which are tremendously important to the ecology and evolution of bacteria, play a key role in development genetic engineering. Bacteriophage virion proteins essential materials infectious viral particles charge several biological functions. The correct identification bacteriophage is great importance for understanding both life at molecular level evolution. However, few computational methods available identifying proteins. In this paper, we proposed new method predict using...

Abstract Halohydrin dehalogenases (HHDHs) possess an unnatural activity of introducing functionalities such as N 3 , CN, NO 2 etc., into a molecule through the ring‐opening reaction epoxides. The enantioselectivity HHDHs is substrate‐dependent and not always high enough for synthetic applications. B‐group has been neglected in past, due to observed low based on performance relatively limited number substrates. Extensive screening substrates HheB2 from Mycobacterium sp. GP1 HheB...

Halohydrin dehalogenase (HheC) from Agrobacterium radiobacter AD1 catalyzes the reversible intramolecular nucleophilic displacement of a halogen by hydroxyl group in vicinal haloalcohols, producing corresponding epoxides. The enzyme displays high enantioselectivity toward some aromatic halohydrins. To understand kinetic mechanism and enzyme, steady-state pre-steady-state analysis was performed with p-nitro-2-bromo-1-phenylethanol (PNSHH) as model substrate. Steady-state analyses indicated...

Halohydrin dehalogenase (HheC) from Agrobacterium radiobacter AD1 catalyzes the dehalogenation of vicinal haloalcohols by an intramolecular substitution reaction, resulting in formation corresponding epoxide, a halide ion, and proton. Halide release is rate-limiting during catalytic cycle conversion (R)-p-nitro-2-bromo-1-phenylethanol enzyme. The recent elucidation X-ray structure HheC showed that hydrogen bonds between OH group Tyr187 Oδ1 atom Asn176 Nε1 Trp249 could play role stabilizing...