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Adipic acid is an industrially important chemical, but the current approach to synthesize it can be of serious pollution to the environment. Rencently, bio-based production of adipic acid has significantly advanced with the development of metabolic engineering and synthetic biology. However, genetic heterogeneity-caused decrease of product titer has largely limited the industrialization of chemicals like adipic acid. Therefore, in the attempt to overcome this challenge, we constitutively expressed the reverse adipate degradation pathway, designed and optimized an adipic acid biosensor, and established a high-throughput screening platform to screen for high-performance strains based on the optimized biosensor. Using this platform, we successfully screened a strain with an adipic acid titer of 188.08 mg·L−1. Coupling the screening platform with fermentation optimization, the titer of adipic acid reached 531.88 mg·L−1 under shake flask fermentation, which achieved an 18.78-fold improvement comparing to the initial strain. Scale-up fermentation in a 5-L fermenter utilizing the screened high-performance strain was eventually conducted, in which the adipic acid titer reached 3.62 g·L−1. Overall, strategies developed in this study proved to be a potentially efficient method in reducing the genetic heterogeneity and was expected to provide guidance in helping to build a more efficient industrial screening process.Key points• Developed a fine-tuned adipic acid biosensor.• Established a high-throughput screening platform to screen high-performance strains.• The titer of adipic acid reached 3.62 g·L−1 in a 5-L fermenter.

Plastic food contact materials (FCM)-based products were widely used in everyday life. These products were normally imposed to strict regulations in order to pass the enforcement tests of compliance as a prefix condition. However, even in these \"qualified\" materials, unknown chemical substances, not involving in legislation lists, could migrate from FCM. In this perspective, the present work aims to thoroughly analyze by means of Gas Chromatography-Mass Spectrometry (GC-MS) the different substances/migrants in 120 qualified FCM plastic products. Unexpectedly, among the identified compounds (nearly 100), only 13% was included in the permitted list of Commission Regulation EU No 10/2011. All the identified compounds were classified into 11 categories according to their chemical structure and the FCM type, whereas toxicology data were in addition analyzed. Each plastic type exhibited different preferences of chemical migrants. Fortunately, most of the compounds identified were of low toxicity, and only 4 chemicals were included in priority lists and previous literature reports as potential risk factors. Subsequently, the accurate amount of these 4 chemicals was determined. The amount of Bis(2-ethylhexyl) adipate (DEHA) and Bis(2-ethylhexyl) phthalate (DEHP) were lower than the SML in Commission Regulation EU No 10/2011, and that of stearamide was under the recommended use quantity. The 2,4-di-tert-butylphenol (2,4-DTBP) was widely exist in the investigated FCM products. Among them, the highest level is obtained in polypropylene/low density polyethylene (BOPP/LDPE) materials, up to 45.568±31.513 mg/kg. In summary, a panel of unlisted chemical migrants were discovered and identified by GS-MS screening. The results implied that plastic FCMs were not so \"inert\" as they usually considered, and further safety evaluation should be performed toward the complete identification of new substances in FCM products.

Abstract Biopolymers based on polylactic acid (PLA) and starch have numerous advantages, such as coming from renewable sources or being compostable, though they can have deficiencies in mechanical properties, and for this reason, polyester resins are occasionally added to them in order to improve their properties. In this work, migration from a PLA sample and from another starch-based biopolymer to three different food simulants was studied. Attention was focused on the determination of oligomers. The analysis was first performed by ultraperformance liquid chromatography quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF–MS), which allowed the identification of the oligomers present in migration. Then, the samples were analyzed by two ambient desorption/ionization techniques directly coupled to mass spectrometry (ADI), direct analysis in real-time coupled to standardized voltage and pressure (DART-MS) and atmospheric pressure solids analysis probe (ASAP-MS). These methodologies were able to detect simultaneously the main oligomers migrants and their adducts in a very rapid and effective way. Nineteen different polyester oligomers, fourteen linear and five cyclic, composed of different combinations of adipic acid [AA], propylene glycol [PG], dipropylene glycol [DPG], 2,2-dibutyl-1,3-propanediol [DBPG], or isobutanol [i-BuOH] were detected in migration samples from PLA. In migration samples from starch-based biopolymer, fourteen oligomers from poly(butylene adipate co-terephthalate) polyester (PBAT) were identified, twelve cyclic and two linear. The results from ADI techniques showed that they are a very promising alternative tool to assess the safety and legal compliance of food packaging materials.

Di-(2-ethylhexyl) adipate (DEHA) and phthalates are commonly used as plasticizers to soften polyvinyl chloride products. Because both DEHA and certain phthalates have been identified as priority chemicals for assessment of human health risk under the Government of Canada's Chemicals Management Plan, a comprehensive targeted survey was conducted to investigate the occurrence of DEHA and eight phthalates (di-methyl phthalate, di-ethyl phthalate, di-n-butyl phthalate, di-iso-butyl phthalate, butyl benzyl phthalate, di-n-hexyl phthalate, d-(2-ethylhexyl) phthalate, and di-n-octyl phthalate) in a total of 118 samples of meat (beef, pork, and chicken), fish, and cheese packaged mostly in cling films. The eight phthalates were not detected in any of the food packaging, but DEHA was detected in most of the cling films, indicating that although DEHA-plasticized films (e.g., polyvinyl chloride film) are currently being used by most grocery stores, nonplasticized cling films such as polyethylene film, are also being used by some stores. DEHA was not detected in any of the 10 cheese samples packaged in nonplasticized rigid plastics but was detected in all 30 cheese samples packaged in DEHA-plasticized cling films at levels from 0.71 to 879 μg/g, with an average of 203 μg/g. Only DEHA was detected in the beef, pork, chicken, and fish samples packaged in DEHA-plasticized cling films but at considerably lower levels than those found in cheese, with averages of 6.3, 9.1, 2.5, and 5.9 μg/g, respectively. Among the eight phthalates, only di-(2-ethylhexyl) phthalate (DEHP) was detected in a few cheese samples at levels from 0.29 to 15 μg/g, with an average of 2.8 μg/g; these levels were very likely due to environmental contamination. Levels of DEHA found in most of the cheese samples from this study are above the European specific migration limit of 18 mg/kg for DEHA in food or food simulants, and levels of phthalates (i.e., DEHP) were low.

Polyvinyl chloride (PVC) food-wrapping films plasticized with di-(2-ethylhexyl) adipate (DEHA) are commonly used by grocery stores in Canada to rewrap meat, poultry, fish, cheese, and other foods. DEHA was assessed as part of the Government of Canada's Chemicals Management Plan. The main source of exposure for most age groups was expected to be food. Although the margin of exposure from food and beverages is considered to be adequately protective, the Government of Canada committed to performing targeted surveys of DEHA in foods and food packaging materials to better define Canadian exposure to DEHA through dietary intake. In order to determine whether more-comprehensive targeted surveys on DEHA in foods should be conducted, 26 food composite samples from the 2011 Canadian total diet study were selected and analyzed for DEHA using a method based on solvent and dispersive solid-phase extraction and gas chromatography–mass spectrometry. These 26 food composites include cheese, meat, poultry, fish, and fast foods, and PVC films were likely used in packaging the individual foods used to make the composites. DEHA was detected in most of the meat, poultry, and fish composite samples, with the highest concentration found in ground beef (11 μg/g), followed by beef steak (9.9 μg/g), freshwater fish (7.8 μg/g), poultry liver pâté (7.4 μg/g), fresh pork (6.9 μg/g), cold cuts and luncheon meats (2.8 μg/g), veal cutlets (2.1 μg/g), roast beef (1.3 μg/g), lamb (1.2 μg/g), and organ meats (0.20 μg/g). Targeted surveys should be conducted to investigate the presence of DEHA in various foods packaged with PVC films in more detail and provide updated occurrence data for accurate human exposure assessment.

Human scent can be collected by either contact or non-contact sampling mode. The most frequently used human scent evidence collection device known as the Scent Transfer Unit (STU-100) is a dynamic sampling device and is often used in a non-contact mode. A customized human scent collection chamber was utilized in combination with controlled odor mimic permeation systems containing five standard human scent volatiles to optimize the flow rate, collection material and geometry of the absorbent material. The scent collection method which yielded the greatest amount of volatile organic compounds (VOCs) detected included the use of a single layer of Johnson and Johnson gauze/multiple layers of Dukal gauze with the STU-100 on the lowest flow rate setting. The correlation of the resulting VOC profiles demonstrate that collection of standard VOCs in controlled conditions yielded reproducible VOC profiles on all materials studied with the exception of polyester. Finally, the method was tested using actual human subjects under optimized set of conditions.

We monitored 26 compounds knowns or suspected to be endocrine disruptors in several environmental water samples from a river, the sea, and an irrigation canal. Because of the various chemical properties of the compounds monitored, analyses were carried out by using two different methods. One method is based on solid‐phase extraction (SPE) on‐line coupled to gas chromatography–mass spectrometry through an on‐column interface. Another is based on high‐performance liquid chromatography–(electrospray) mass spectrometry working in negative ionization mode and using off‐line SPE. The limits of detection for the two methods were at levels of low μg/L. Phthalates and bis‐(2‐ethylhexyl) adipate were found at levels between 0.05 and 13 μg/L in all of the water samples analyzed. Some pesticides, alkylphenols, and estrogens were determined in a few samples at levels below 0.1 μg/L.

Pseudomonas aeruginosa, isolated from soil near tannery effluent was able to degrade 8-anilino-1-naphthalenesulfonic acid (ANSA), a sulfonated aromatic amine. The organism degraded this amine up to a concentration of 1,200 mg l-1 using glucose and ammonium nitrate as carbon and nitrogen sources respectively. The degradation started when the organism reached its late exponential growth phase. Salicylic acid and β-ketoadipic acid were identified as intermediate compounds using HPLC and GC-MS and provide evidence for ortho pathway reactions. Further proof for the pathway is obtained from the dioxygenase activity of the strain growing exponentially in medium with ANSA and glucose.

Acinetobacter lwoffii K24 is a soil bacterium that can use aniline as a sole carbon and nitrogen source (by beta-ketoadipate pathway genes (cat genes)) and has two copies of catABC gene separately located on the chromosome. In order to identify aniline-induced proteins, two-dimensional electrophoresis (2-DE) was applied to soluble protein fractions of A. lwoffii K24 cultured in aniline and succinate media. In the range of pH3-10, more than 370 spots were detected on the silver stained gels. Interestingly, more than 20 spots were selectively induced on aniline-cultured bacteria. Twenty-three protein spots of A. lwoffii K24 were analyzed by N-terminal microsequencing and internal microsequencing with in-gel digestion. Of 20 aniline induced protein spots, we identified six beta-ketoadipate pathway genes, one subunit of amino group transfer (putative subunit of aniline oxygenase), malate dehydrogenase, putative ABC transporter, putative hydrolase, HHDD isomerase, and five unknown proteins. Especially in case of two catechol 1,2-dioxygenases (CDI1 and CDI2), more than three isotypes were detected on the 2D gel. This study showed that the proteome analysis of A. lwoffii K24 may be helpful for identification of genes induced by aniline and understanding of their function in the cell.

Background Biobased processes for the production of adipic acid are of great interest to replace the current environmentally detrimental petrochemical production route. No efficient natural producer of adipic acid has yet been identified, but several approaches for pathway engineering have been established. Research has demonstrated that the microbial production of adipic acid is possible, but the yields and titres achieved so far are inadequate for commercialisation. A plausible explanation may be intolerance to adipic acid. Therefore, in this study, selected microorganisms, including yeasts, filamentous fungi and bacteria, typically used in microbial cell factories were considered to evaluate their tolerance to adipic acid. Results Screening of yeasts and bacteria for tolerance to adipic acid was performed in microtitre plates, and in agar plates for A. niger in the presence of adipic acid over a broad range of concentration (0–684 mM). As the different dissociation state(s) of adipic acid may influence cells differently, cultivations were performed with at least two pH values. Yeasts and A. niger were found to tolerate substantially higher concentrations of adipic acid than bacteria, and were less affected by the undissociated form of adipic acid than bacteria. The yeast exhibiting the highest tolerance to adipic acid was Candida viswanathii , showing a reduction in maximum specific growth rate of no more than 10–15% at the highest concentration of adipic acid tested and the tolerance was not dependent on the dissociation state of the adipic acid. Conclusions Tolerance to adipic acid was found to be substantially higher among yeasts and A. niger than bacteria. The explanation of the differences in adipic acid tolerance between the microorganisms investigated are likely related to fundamental differences in their physiology and metabolism. Among the yeasts investigated, C. viswanathii showed the highest tolerance and could be a potential host for a future microbial cell factory for adipic acid.