Metabolic engineering of Escherichia coli for adipic acid production / 生物工程学报

Adipic acid is a high-value-added dicarboxylic acid which is primarily used in the production of nylon-66 for manufacturing polyurethane foam and polyester resins. At present, the biosynthesis of adipic acid is hampered by its low production efficiency. By introducing the key enzymes of adipic acid reverse degradation pathway into a succinic acid overproducing strain Escherichia coli FMME N-2, an engineered E. coli JL00 capable of producing 0.34 g/L adipic acid was constructed. Subsequently, the expression level of the rate-limiting enzyme was optimized and the adipic acid titer in shake-flask fermentation increased to 0.87 g/L. Moreover, the supply of precursors was balanced by a combinatorial strategy consisting of deletion of sucD, over-expression of acs, and mutation of lpd, and the adipic acid titer of the resulting E. coli JL12 increased to 1.51 g/L. Finally, the fermentation process was optimized in a 5 L fermenter. After 72 h fed-batch fermentation, adipic acid titer reached 22.3 g/L with a yield of 0.25 g/g and a productivity of 0.31 g/(L·h). This work may serve as a technical reference for the biosynthesis of various dicarboxylic acids.

Application of cutinase in the degradation of biodegradable polyester poly(butylene adipate-co-terephthalate) / 生物工程学报

With the environmental pollution caused by waste plastics becoming increasingly serious, biodegradable polyester has become the focus of public attention. Poly(butylene adipate-co-terephthalate) (PBAT) is a biodegradable polyester formed by the copolymerization of aliphatic and aromatic groups, which has excellent performance of both. The degradation of PBAT under natural conditions requires strict environmental conditions and long degradation cycle. To address these shortcomings, this study explored the application of cutinase in PBAT degradation and the impact of butylene terephthalate (BT) content on the biodegradability of PBAT, so as to improve the degradation rate of PBAT. Five Polyester degrading enzymes from different sources were selected to degrade PBAT to pick out the most efficient enzyme. Subsequently, the degradation rate of PBAT materials with different BT content were determined and compared. The results showed that cutinase ICCG was the best enzyme for PBAT biodegradation, and the higher the BT content, the lower the degradation rate of PBAT. Furthermore, the optimum temperature, buffer type, pH, the ratio of enzyme to substrate (E/S) and substrate concentration in the degradation system were determined to be 75 ℃, Tris HCl, 9.0, 0.4% and 1.0% respectively. These findings may facilitate the application of cutinase in PBAT degradation.

Synthesis of diisooctyl adipate catalyzed by lipase-displaying Pichia pastoris whole-cell biocatalysts / 生物工程学报

An enzyme-displaying yeast as a whole-cell biocatalyst is an alternative to immobilized enzyme, due to its low-cost preparation and simple recycle course. Here, lipase-displaying Pichia pastoris whole-cell was used as a biocatalyst to synthesize diisooctyl adipate in the non-aqueous system. The maximum productivity of diisooctyl adipate was obtained as 85.0% in a 10 mL reaction system. The yield could be reached as high as 97.8% when the reaction system was scaled up to 200 mL. The purity obtained is 98.2% after vacuum distillation. Thus, the lipase-displaying P. pastoris whole-cell biocatalyst was promising in commercial application for diisooctyl adipate synthesis in non-aqueous phase.

Preface for special issue on biobased chemicals (2013) / 生物工程学报

Biobased chemicals are one of the main missions of bioeconomy. In this special issue, we reviewed the recent progress in the metabolic engineering and fermentation control study on biobased succinic acid, adipic acid, lactic acid, 3-hydroxypropanoic acid, glucaric acid, glycerol, xylitol, higher alcohols and ethylene, recombinant construction for the direct utilization of lignocelluloses, biotransformation of bio-based lactic acid, and salting-out extraction of bio-based chemicals. Some research articles on biobased succinic acid, D-mannitol, malic acid, 5-aminolevulinic acid, 1,3-propanediol, and butanol are also included.

Biosynthesis of adipic acid / 生物工程学报

Adipic acid is a six-carbon dicarboxylic acid, mainly for the production of polymers such as nylon, chemical fiber and engineering plastics. Its annual demand is close to 3 million tons worldwide. Currently, the industrial production of adipic acid is based on the oxidation of aromatics from non-renewable petroleum resources by chemo-catalytic processes. It is heavily polluted and unsustainable, and the possible alternative method for adipic acid production should be developed. In the past years, with the development of synthetic biology and metabolic engineering, green and clean biotechnological methods for adipic acid production attracted more attention. In this study, the research advances of adipic acid and its precursor production are reviewed, followed by addressing the perspective of the possible new pathways for adipic acid production.

Metabonomic profiling of plasma metabolites in Wistar rats to study the effect of aging by means of GC/TOFMS-based techniques / 药学学报

The global metabolite profiles of endogenous compounds of Wistar rats from 12 to 20 weeks old were investigated to take deep insight into and get better understanding of the pathogenesis of development and aging. Plasma from Wistar rats at 12, 14, 16, 18, and 20 weeks old were analyzed using GC/TOFMS. Multivariate data analysis was then used to process the metabonomic data which indicated excellent separation between different weeks and showed that the metabolic profiles of the samples changed with age, enabling age-related metabolic trajectories to be visualized. Decreased concentrations of citric acid, cis-aconitic acid, 9-(z)-hexadecenoic acid along with increased levels of hexanedioic acid, alpha-tocopherol, 3-indole propionic acid, etc contributed to the separation. Several major metabolic pathways were identified to be involved in metabolic regulation. This suggests that GC/TOFMS-based metabonomics is a powerful alternative approach to identifying potential biomarkers and investigating the physiological developments of aging and it is important to employ suitable age-match control group in metabonomic study of physiological monitoring, drug safety assessment, and disease diagnosis, etc.

The effect of thyroidectomy on male rats fed two antagonist lipid diets. IV. The fatty acid content of mitochondrial lipids of heart and gastrocnemic muscles

The fatty acid (FA) contents of heart and gatrocnemic muscle mitochondrial lipids of thyroidecromized and control (sham-operated) were analyzed. Animals were fed either an n-6 polyunsaturated (PUFA)-rich diet A or an n- PUFA-rich diet B during 30 days. It was observed alteraions in the composition of FA of those cell organelles caused by the absence of the thyroid gland. Some of these alterations were dependent of the type of used diet. Thyroidecromy caused an increase of P?S ratio in heart and gastrocnemic muscle but only for animals fed diet B. Thyroidectomy caused an increase of n-6/n-3 ratio in heart of rats fed diet A and in gastrocnemic muscle of rats fed diet B and it caused a decrease of n-6/n-3 ratio of heart of rats fed diet A and in gastrocnemic muscle of rats fed diet B. The absence of thyroid gland in some way may interfer in the FA metabolism and in the rate of FA incorporation into endogeneous lipids but in a different process dependent of type of tissue and of used lipid diet

Evaluación comparativa entre ácido kaínic, santonina y adipato de piperazina solos y asociados como antihelmínticos: terapétuica y tolerancia / Comparative evaluation between kainic acid, santonine and piperazine adipate alone and combined as antihelminthes: therapeutics and tolerance

Se estudió a la población escolar de una zona endémica para geohelmintiasis mediante estudios coproparasitoscópicos, utilizando el método de flotación. Se seleccionaron con 200 pacientes que presentaban parasitosis por Ascaris lumbricoides, Trichuris trichiura y uncinaria. En un estudio posterior se valoraron por el método de frotis fecal de Kato Katz, además de realizar tres estudios por el método de raspado anal con cinta de celulosa engomada. Los pacientes se dividieron en cinco grupos y se les administró en una sola dosis: grupo 1, santonina a concentración de 66.6 mg/10 ml; grupo 2, adipato de piperazina a concentración de 1333.2 mg/10 ml, grupo 3, ácido Kaínico a concentración de 6.6 mg/10 ml; grupo 4, combinación de santonina 33.3 mg/10 ml, piperazina 666.6 mg/10 ml y ácido Kaínico 3.3 mg/10 ml, y grupo 5, adipato de piperazina a concentración de 2666.4 mg/10ml. Posterior al tratamiento se practicaron nuevamente los exámenes mencionados a manera de control. Los resultados demostraron que la combinación de ácido Kaínico, piperazina y santonina es mejor en el control de la helmintiasis que los fármacos utilizados en forma independiente