BACKGROUND: Degradation reactions of micropollutants such as antibiotics with OH radicals are very important in terms of environmental pollution. Therefore, in this study, the degradation kinetic mechanism of 6-aminopenicillanic acid (6-APA) with OH radical was investigated by density functional theory (DFT) methods.
METHODS: For the calculations, different functionals such as B3LYP, MPW1PW91, and M06-2X were used with a 6-31 g(d,p) basis set. The aquatic effect on the reaction mechanism was investigated by conductor-like polarizable continuum model (CPCM). For the degradation kinetics in aqueous media, the addition of explicit water molecules was also calculated. Subsequent reaction mechanism for the most probable reaction product was briefly discussed.
RESULTS: Among the functionals used, B3LYP results were consistent with the experimental results. Calculated kinetic parameters indicated that the OH-addition path was more dominant than the H-abstraction paths. With the increase of explicit water molecules in the models, the energy required for the formation of transition state complexes decreased. The overall rate constant is calculated as 2.28 × 1011 M-1 s-1 at 298 K for the titled reaction.

The present study deals with developing vermiculite (VMT)-alginate (Alg) composites with different cross-linker concentrations (CaCl2) to deliver the controlled 6-aminopenicillin acid (6-APA). The Characterization of synthesized composites was conducted by Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analyses. Optimization attempts were explored via the response surface method (RSM) to best predict the actual amount of compound. The adsorption capacity of 6-APA onto this adsorbent was found to be 208.33 mg/g, which was higher than that for other clays. The equilibrium and Kinetic studies (chemical reaction and diffusion-based models) indicated that drug absorption on VMT-Alg is homogeneous with chemical interaction. An increase in cross-linker (CaCl2) concentration leads to improvement in the drug encapsulation efficiency while having no significant effect on loading efficiency. The in-vitro release of the pure drug shows a rapid burst release followed by 100% cumulative release within 6 h. Whereas, the synthesized drug with Alg substantially showed less release of 43% after 8 h. Release experiments revealed that the presence of the CaCl2 delayed the release of the 6-APA less than 35% after 12 h. The kinetic release of 6-APA is followed by the Korsmeyer-Peppas model based on Fick\'s law mechanism due to the kinetic exponent (n < 0.5). All studied composites antibacterial activity after 24 h exposure against E. Coli and S. aureus. The antibacterial activities of composites were evaluated by the halo of no growth. The results showed that the VMT-Alg-6APA composite had strong activity against Gram-positive and Gram-negative bacteria.

BACKGROUND: Penicillin G amidase/acylases from microbial sources is a unique enzyme that belongs to the N-terminal nucleophilic hydrolase structural superfamily. It catalyzes the selective hydrolysis of side chain amide/acyl bond of penicillins and cephalosporins whereas the labile amide/acyl bond in the β-lactam ring remains intact. This review summarizes the production aspects of PGA from various microbial sources at optimized conditions. The minimal yield from wild strains has been extensively improved using varying strain improvement techniques like recombination and mutagenesis; further applied for the subsequent synthesis of 6-aminopenicillanic acid, which is an intermediate molecule for synthesis of a wide range of novel β-lactam antibiotics. Immobilization of PGA has also been attempted to enhance the durability of enzyme for the industrial purposes.
CONCLUSIONS: The present review provides an emphasis on exploitation of E. coli to enhance the microbial production of PGA. The latest achievements in the production of recombinant enzymes have also been discussed. Besides E. coli, other potent microbial strains with PGA activity must be explored to enhance the yields.

Processes employed in separations of products of enzyme reactions are often driven by diffusion, and their efficiency can be limited. Here, we exploit the effect of a direct current (DC) electric field that intensifies mass transfer through a semipermeable membrane for fast, continuous, and selective separation of electrically charged molecules. Specifically, we separate low-molecular-weight reaction products (phenylacetic acid, 6-aminopenicillanic acid) from the original reaction mixture containing a free enzyme (penicillin acylase). The developed microfluidic dialysis-membrane contactor allows a stable counter-current arrangement of the retentate and permeates liquid streams on which DC electric field is perpendicularly applied. The applied electric field significantly accelerates the transport of electrically charged products through the semipermeable membrane yielding high separation efficiencies at short residence times. The residence time of 5 min is sufficient to reach 100% separation yield in the electric field. The same residence time provides only a 50% yield in the diffusion-controlled experiments. We experimentally demonstrated that a combined microreactor-microextractor with a recycle of the soluble penicillin acylase can continuously produce both the reaction products at high concentrations. The developed membrane-contactor is a versatile platform allowing to tune its characteristics, such as selectivity given by the membrane, or the type of the retentate phase, for a specific application.

Aqueous two-phase systems (ATPSs) were screened for the production of 6-aminopenicillanic acid (6-APA) catalyzed by penicillin acylase, followed by the extractive separation of 6-APA from the reaction mixture. The key point of this study was to find an ATPS exhibiting a large difference in the partition coefficients of the biocatalyst and reaction products. Several ATPSs based on polyethylene glycol (PEG)/phosphate, PEG/citrate, and PEG/dextran were tested. We found that an ATPS consisting of 15 wt% of PEG 4000, 10 wt% of phosphates, 75 wt% of water (pH value 8.0 after dissolution) provided optimal separation of 6-APA from the enzyme. While the 6-APA was mainly found in the top PEG phase, the free enzyme favored the bottom salt-rich phase. This ATPS also fulfils other important requirements: (i) high buffering capacity, reducing an undesirable pH decrease due to the dissociation of phenylacetic acid (the side product of the reaction), (ii) a relatively low cost of the ATPS components, (iii) the possibility of electrophoretic transport of fine droplets as well as the reaction products for both the acceleration of phase separation and the enhancement of 6-APA concentration in the product stream. Extraction experiments in microcapillary and batch systems showed that the transport of 6-APA formed in the salt-rich phase to the corresponding PEG phase could occur within 30 s. The experimental results described form a base of knowledge for the development of continuously operating integrated microfluidic reactors-separators driven by an electric field for the efficient production of 6-APA.

Objective: To observe serum levels of periostin, ECP, IgE in the antibiotic enterprise workers, and study the role of periostin, ECP, IgE in the development of allergic inflammation. Methods: 90 cases with asthma or rhinitis were enrolled as disease group, another 117 workers exposed to 7-ACA、6-APA dust without suffering from allergic illness, are chosen as group of dust exposed, and 192 healthy workers who didn\'t contact dust were chosen as control group. Questionnaires were used to learn their basic information.Lung function was determined with a portable spirometer.The expression levels of periostin、ECP and IgE in serum were measured by enzyme-linked immuno sorbent assay. Results: The exposure group and disease group had significantly lower forced vital capacity (FVC) , forced expiratory volume in 1 second (FEV(l.0)) , and FEV(l.0)/FVC ratio than the control group (P<0.05) . The disease group had significantly higher eosinophil than the control group (P<0.05) . Compared with the control group, the exposure group, the disease group, asthma subgroup, rhinitis subgroup of serum periostin and IgE increased, the differences are statistically significant (P<0.05) . Serum levels of ECP in the workers of asthma subgroup were significantly higher than that in control group (P<0.05) . Serum expression levels of periostin were positively correlated with IgE, ECP in workers (P<0.001) , serum levels of periostin were negatively correlated with FEV(1.0) in workers (P<0.05) . Multiple logistics regression analysis found that exposure to 7-ACA or 6-APA (OR=3.09, 95%CI: 1.83-5.21) , age>47years (OR=2.53, 95%CI: 1.22-5.26) , higher ECP (OR=1.04, 95%CI: 1.01-1.06) were risk factors for increased serum periostin level. Conclusion: Occupational exposure to 7-ACA or 6-APA can result in higher serum periostin level, exposure to 7-ACA or 6-APA, age>47 years, higher ECP are risk factors for increased serum periostin level.
目的： 观察β-内酰胺类抗生素接触工人血清骨膜蛋白（periostin）、嗜酸粒细胞阳离子蛋白（ECP）、免疫球蛋白E（IgE）水平，探讨其在过敏性炎症发生发展中的作用。 方法： 选取90名接触7-氨基头孢烷酸（7-ACA）、6-氨基青霉烷酸（6-APA）粉尘哮喘或过敏性鼻炎患者为病例组（哮喘亚组32人，鼻炎亚组58人），选取117名接触7-ACA、6-APA粉尘无哮喘或鼻炎等过敏性疾病工人为接触组，选择192名不接触粉尘体检健康者作为对照组。通过问卷调查基本情况，便携式肺功能仪测定肺功能，ELISA法测定血清骨膜蛋白、ECP和IgE水平。 结果： 接触组和病例组工人用力肺活量（FVC）、第1秒用力呼气容积（FEV(1.0)）及FEV(1.0)/FVC均低于对照组，差异有统计学意义（P<0.05）。病例组嗜酸粒细胞计数明显高于对照组，差异有统计学意义（P<0.05）。接触组、病例组、哮喘亚组、鼻炎亚组血清骨膜蛋白、IgE水平均高于对照组，哮喘亚组血清ECP水平高于对照组，差异有统计学意义（P<0.05）。血清骨膜蛋白与IgE、ECP均呈正相关（P<0.01），血清骨膜蛋白与FEV(1.0)呈负相关（P<0.05）。多元logistic回归分析发现，接触7-ACA或6-APA（OR=3.09，95%CI：1.83~5.21）、年龄>47岁（OR=2.53，95%CI：1.22~5.26）、ECP增高（OR=1.04，95%CI：1.01~1.06）为血清骨膜蛋白升高的危险因素。 结论： 职业接触7-ACA、6-APA可导致血清骨膜蛋白表达水平升高，接触7-ACA或6-APA、年龄>47岁、ECP增高可作为骨膜蛋白升高的危险因素。.

Ethyl 4-amino-2-fluorophenylpiperazin-1-carboxylates containing a 1,3-oxazol(idin)e, 5-thioxo-1,2,4-triazole, 1,3,4-thiadiazole, 5-thioxo-1,3,4-oxadiazole, or 1,3-thiazole nucleus were obtained starting from ethyl piperazine-1-carboxylate (1) by several steps. The treatment of amine, 3 or hydrazide, 9 with several aromatic aldehydes generated the corresponding arylmethyleneamino (3a-f) or arylidenehydrazino (12a-c) compounds. The Mannich reaction between the 1,2,4-triazole or 1,3,4-oxadiazole compounds and 7-aca produced cephalosporanic acid derivatives. Penicillanic acid derivatives were obtained when 6-apa was used in the Mannich reactions. The synthesized compounds were screened for their antimicrobial, antilipase, and antiurease activities. Some of them were found to possess good-moderate antimicrobial activity against the test microorganisms. Two compounds exhibited antiurease activity, and four of them displayed antilipase activity.