Chlorophenols are environmental contaminants, which are highly toxic to living beings due to their carcinogenic, mutagenic and cytotoxic properties. Bacterial degradation has been considered a cost-effective and eco-friendly method of removing chlorophenols, compared to the traditional physical–chemical processes. In this study, we first developed an efficient process for the biodegradation of chlorophenols by magnetically immobilized Rhodococcus rhodochrous cells. R. rhodochrous DSM6263 degrades chlorophenols following the first step of hydroxylation at the ortho-positions of chlorophenolic rings. The cells immobilized by k-carrageenan with 9 g/L Fe3O4 nanoparticles could efficiently degrade 2-chlorophenol, 4-chlorophenol, 2,3-dichlorophenol and their mixture, which were even higher than those by free cells. The magnetically nanoparticle-immobilized cells could be used at least for six cycles. Given the much easier separation by an external magnetic field and high degradation efficiency, this study provides a promising technique for improving biocatalysts used in the bioremediation process for chlorophenols in wastewater.

Metopolophium dirhodum(Walker) (Hemiptera: Aphididae) is one of the most common aphid pests of winter cereals. To facilitate accurate gene expression analyses with qRT-PCR assays, the expression stability of candidate reference genes under specific experimental conditions must be verified before they can be used to normalize target gene expression levels. In this study, 10 candidate reference genes inM.dirhodumwere analyzed by qRT-PCR under various experimental conditions. Their expression stability was evaluated with delta Ct, BestKeeper, geNorm, and NormFinder methods, and the final stability ranking was determined with RefFinder. The results indicate that the most appropriate sets of internal controls wereSDHBandRPL8across geographic population;RPL8,Actin, andGAPDHacross developmental stage;SDHBandNADHacross body part;RPL8andActinacross wing dimorphism and temperature;RPL4andEF1Aacross starvation stress;AKandRPL4across insecticide treatments;RPL8andNADHacross antibiotic treatments;RPL8,RPL4,Actin, andNADHacross all samples. The results of this study provide useful insights for establishing a standardized qRT-PCR procedure forM. dirhodumand may be relevant for identifying appropriate reference genes for molecular analyses of related insects.

Alkaline pectate lyases have biotechnological applications in plant fiber processing, such as ramie degumming. Previously, we characterized an alkaline pectate lyase from Bacillus clausii S10, named BacPelA, which showed potential for enzymatic ramie degumming because of its high cleavage activity toward methylated pectins in alkaline conditions. However, BacPelA displayed poor thermo-alkaline stability. Here, we report the 1.78 Å resolution crystal structure of BacPelA in apo form. The enzyme has the characteristic right-handed β-helix fold of members of the polysaccharide lyase 1 family and shows overall structural similarity to them, but it displays some differences in the details of the secondary structure and Ca²⁺-binding site. On the basis of the structure, 10 sites located in flexible regions and showing high B-factor and positive ΔT_m values were selected for mutation, aiming to improve the thermo-alkaline stability of the enzyme. Following site-directed saturation mutagenesis and screening, mutants A238C, R150G, and R216H showed an increase in the T₅₀¹⁵ value at pH 10.0 of 3.0 °C, 6.5 °C, and 7.0 °C, respectively, compared with the wild-type enzyme, interestingly accompanied by a 24.5%, 46.6%, and 61.9% increase in activity. The combined mutant R150G/R216H/A238C showed an 8.5 °C increase in the T₅₀¹⁵ value at pH 10.0, and an 86.1% increase in the specific activity at 60 °C, with approximately doubled catalytic efficiency, compared with the wild-type enzyme. Moreover, this mutant retained 86.2% activity after incubation in ramie degumming conditions (4 h, 60 °C, pH 10.0), compared with only 3.4% for wild-type BacPelA. The combined mutant increased the weight loss of ramie fibers in degumming by 30.2% compared with wild-type BacPelA. This work provides a thermo-alkaline stable, highly active pectate lyase with great potential for application in the textile industry, and also illustrates an effective strategy for rational design and improvement of pectate lyases.

Lactic acid has been identified as one of the top 30 potential building-block chemicals from biomass. Therefore, the search for cheap raw materials is an objective to reduce the production costs. Efficient polymer-grade L-lactic acid production was achieved in this report by a thermophilic strain Bacillus sp. XZL4 using corn stover hydrolyzate as sole carbon source. High L-lactic acid concentration (81.0 g L^-1) was obtained from 162.5 g L^-1 concentrated corn stover hydrolyzate (total reducing sugar of 83.0 g L^-1) with a volumetric productivity of 1.86 g L^-1 h^-1 (0–36 h) and a product yield of 0.98 g g^-1 total reducing sugars. This is the highest L-lactic acid concentration and yield reported from corn stover hydrolyzate. And the high optical purity of L-lactic acid obtained in this study also indicated that Bacillus sp. XZL4 is a promising polymer-grade L-lactic-acid producer from cellulosic biomass.