Yanhua Hou

The enzyme catechol 1,2‐dioxygenase (CAT) plays a critical role in the biosynthesis pathway of cis, cis‐muconic acid (CCMA), which serves as an indispensable raw material for various industrial applications. In this research, we cloned a novel cold‐adapted CAT (HaCAT) from the Antarctic sea ice bacterium Halomonas sp. ANT108. Homology modeling analysis revealed that HaCAT possessed the characteristic Fe³⁺ binding site and catalytic active site of typical CATs, and it exhibited unique structural adaptations to cold environments. The optimal temperature and pH for recombinant HaCAT (rHaCAT) were found to be 25°C and 6.5, respectively. At 0°C, the enzyme retained a maximum activity of 43.6%, and in the presence of 1.0 M NaCl, its activity reached 173.9%, demonstrating significant salt tolerance. Additionally, the V_max and K_m of rHaCAT were 6.68 μmol/min/mg and 128.90 μM at 25°C, respectively. Furthermore, rHaCAT was successfully immobilized in the metal‐organic framework ZIF‐8 and retained almost 50% of its activity after five reuse cycles, demonstrating excellent reusability. Overall, these results provided a new resource and theoretical foundation for the industrial biocatalytic production and modification of CAT.

Aliphatic nitro compounds cause environmental pollution by being discharged into water with industrial waste. Biodegradation needs to be further explored as a green and pollution-free method of environmental remediation. In this study, we successfully cloned a novel nitronate monooxygenase gene (psnmo) from the genomic DNA library of Psychrobacter sp. ANT206 and investigated its ability to degrade 2-nitropropane (2-NP). Homology modeling demonstrated that PsNMO had a typical I nitronate monooxygenase catalytic site and cold-adapted structural features, such as few hydrogen bonds. The specific activity of purified recombinant PsNMO (rPsNMO) was 97.34 U/mg, rPsNMO exhibited thermal instability and reached maximum catalytic activity at 30 °C. Moreover, rPsNMO was most active in 1.5 M NaCl and remained at 104% of its full activity in 4.0 M NaCl, demonstrating its significant salt tolerance. Based on this finding, a novel bacterial cold-adapted enzyme was obtained in this work. Furthermore, rPsNMO protected E. coli BL21 (DE3)/pET28a(+) from the toxic effects of 2-NP at 30 °C because the 2-NP degradation rate reached 96.1% at 3 h and the final product was acetone. These results provide a reliable theoretical basis for the low-temperature degradation of 2-NP by NMO.

Serine hydroxymethyltransferase (SHMT) plays a significant role in the synthesis of l‐serine, purine, and thymidylate, which could be extensively applied in the treatment of cancers and the development of antibiotics. In this study, cloned from Psychrobacter sp. ANT206, a novel cold‐adapted SHMT gene (psshmt, 1257 bp) encoding a protein of 418 amino acids was expressed in Escherichia coli. The homology modeling result revealed that PsSHMT owned fewer Proline (Pro) residues and hydrogen bonds compared with its homologs from mesophilic E. coli and thermophilic Geobacillus stearothermophilus. In addition, the molecular weight of the purified recombinant PsSHMT (rPsSHMT) was identified to be 45 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis, approximately. The enzymatic characteristics of the cold‐adapted rPsSHMT displayed that its optimum temperature and pH were 30°C and 7.5, respectively, and its enzymatic activity could be inhibited by Cu²⁺, significantly. rPsSHMT also showed a high k_cat value and low ΔG at low temperatures. Furthermore, arginine (Arg) could affect the activity of rPsSHMT and be vital to its active sites. The results of this study reflected that these characteristics of the cold‐adapted rPsSHMT made it a remarkable candidate that could be utilized in multiple industrial fields under low temperatures.

As a global environmental pollution problem, heavy metal pollution has brought great harm to human beings. In this work, we studied the toxicity of Hg²⁺ on allophycocyanin (APC) at the molecular level. Firstly, APC was extracted and purified from Spirulina platensis mud and its purity (A₆₅₀/A₂₈₀) reached 3.75. In addition, the fluorescence intensity of APC decreased with increasing Hg²⁺ concentration from 0 to 5 × 10^-6 mol L⁻¹. The theoretical calculation and experimental results showed that the fluorescence quenching of APC by Hg²⁺ was static and had a good linear relationship. Moreover, the UV-Vis spectra of APC showed a significant decrease at 200 nm and 650 nm with the increase of Hg²⁺ concentration from 0 to 5×10⁻⁶ mol L⁻¹, and a red-shift at 200 nm, which indicated that Hg²⁺ not only affected the structure of APC but also affected the light absorption and photosynthetic function of APC. Furthermore, the results of molecular simulation demonstrate that Hg²⁺ combinations with the Met77, Cys81 in the α chain and the Arg77, Cys81 in the β chain, which interact between the peptide chain and the chromophore, and Hg²⁺ forms a Hg-S bond with -SH. This study provides new insights into the structure and how Hg²⁺ effect the optical properties of APC.

In this study, R-phycoerythrin (R-PE) was isolated and characterized from Porphyra yezoensis by three-phase partitioning (TPP) method. The effects of temperature, time, pH, salt saturation, and volume ratio on the purity and recovery rate of R-PE were studied. The optimum extraction conditions were determined as follows: salt saturation of 70%, temperature of 25 °C, time of 45 min, pH of 7.0, and volume ratio of 1:1. Under the optimal extraction conditions, the purity of R-PE was 3.90. The results of SDS-PAGE showed that R-PE has three bands at 23 kDa, 22 kDa, and 18 kDa, corresponding to its α, β, γ subunits. The structure and optical activity of R-PE did not change before and after purification based on ultraviolet, infrared, and fluorescence spectra. In addition, the purity and recovery rate of R-PE extracted by tert-butanol were evaluated. The results showed that the extraction performance of tert-butanol for R-PE remained unchanged in three recoveries. These show that TPP is an efficient, green, and recyclable extraction technology.

Efficient biodegradation may offer a solution for the treatment of nitro-aromatic compounds (NACs) with toxicity, mutagenicity and persistence in the environment. In this study, dopamine (DA) functionalized magnetic nanoparticles with biocompatibility and hydrophilicity were synthesized and utilized for the immobilization of nitro-aromatic compounds degrading psychrophile Psychrobacter sp. ANT206 harboring the cold-adapted nitroreductase. The prepared nanocarriers were characterized using multiple methods. The highest immobilization yield of cells immobilized by Fe₃O₄@SiO₂@DA was 90.67% under the optimized conditions of 10 °C, pH 7.5, 2 h and cell/support 1.2 mg/mg, and the activity recovery was 89.41%. In addition, the obtained immobilized cells displayed excellent salinity stability and reusability. Moreover, immobilized P. sp. ANT206 strains showed remarkable biodegradation capability on nitrobenzene and p-nitrophenol. This study introduced those novel Fe₃O₄@SiO₂@DA nanoparticles could be applied as ideal and low-cost nanocarriers for the immobilization of cells and large-scale bioremediation of hazardous NACs with perspective applications under low temperature.