Analysis of the microbial community structure and flavor components succession during salt-reducing pickling process of zhacai (preserved mustard tuber).

The salt-reducing pickling method has been applied to the industrial production of zhacai. In order to reveal the succession of the microbial community structure and flavor components during the pickling process, this study used PacBio Sequel to sequence the full length of 16S rRNA (bacteria, 1400 bp) and ITS (fungi, 1200 bp) genes, and detected flavor components simultaneously, including organic acids, volatile flavor components (VFC), monosaccharides, and amino acids. Eleven phyla and 148 genera were identified in the bacterial community, and 2 phyla and 60 genera in the fungal community. During the four stages of pickling, the dominant bacterial genera were Leuconostoc, Lactobacillus, Leuconostoc, and Lactobacillus, while the dominant fungal genera were Aspergillus, Kazachstania, Debaryomyces, and Debaryomyces, respectively. There were 32 main flavor components (5 organic acids, 19 VFCs, 3 monosaccharides, and 5 amino acids). Correlation heat mapping and bidirectional orthogonal partial least squares (O2PLS) analysis showed that the flora having close relation to flavor components included 14 genera of bacteria (Leuconostoc, Clostridium, Devosia, Lactococcus, Pectobacterium, Sphingobacterium, Serratia, Stenotrophomonas, Halanaerobium, Tetragenococcus, Chromohalobacter, Klebsiella, Acidovorax, and Acinetobacter) and 3 genera of fungi (Filobasidium, Malassezia, and Aspergillus). This study provides detailed data regarding the microbial community and flavor components during the salt-reducing pickling process of zhacai, which can be used as a reference for the development and improvement of salt-reducing pickling methods.

Expanding the application of tryptophan: Industrial biomanufacturing of tryptophan derivatives.

Tryptophan derivatives are various aromatic compounds produced in the tryptophan metabolic pathway, such as 5-hydroxytryptophan, 5-hydroxytryptamine, melatonin, 7-chloro-tryptophan, 7-bromo-tryptophan, indigo, indirubin, indole-3-acetic acid, violamycin, and dexoyviolacein. They have high added value, widely used in chemical, food, polymer and pharmaceutical industry and play an important role in treating diseases and improving life. At present, most tryptophan derivatives are synthesized by biosynthesis. The biosynthesis method is to combine metabolic engineering with synthetic biology and system biology, and use the tryptophan biosynthesis pathway of Escherichia coli, Corynebacterium glutamicum and other related microorganisms to reconstruct the artificial biosynthesis pathway, and then produce various tryptophan derivatives. In this paper, the characteristics, applications and specific biosynthetic pathways and methods of these derivatives were reviewed, and some strategies to increase the yield of derivatives and reduce the production cost on the basis of biosynthesis were introduced in order to make some contributions to the development of tryptophan derivatives biosynthesis industry.

Effects of Different Low-Temperature Storage Methods on the Quality and Processing Characteristics of Fresh Beef.

Low-temperature storage has become the most common way for fresh meat storage because of its lower cost and better preservation effect. Traditional low-temperature preservation includes frozen storage and refrigeration storage. The refrigeration storage has a good fresh-keeping effect, but the shelf life is short. Frozen storage has a long shelf life, but it has a great impact on the quality of meat structure and other qualities, and cannot achieve a complete "fresh-keeping" effect. With the development of food processing storage and freezing technology, two new storage methods, ice temperature storage and micro-frozen storage, have attracted more attention. In this paper, the effects of different low-temperature storage methods on the sensory, physicochemical properties, myofibrillar protein oxidation, microstructure, and processing characteristics of fresh beef were studied. The optimal storage methods under different storage requirements were analyzed to reveal the mechanism and efficacy of ice temperature storage and micro-frozen storage technology, as well as the advantages compared with traditional low-temperature refrigeration. It has practical significance for guiding the application of low-temperature storage of fresh meat. Finally, this study concluded that the longest shelf life could be achieved by frozen storage, and the best preservation effect was achieved during the shelf life of ice temperature storage, and the effect of micro-frozen storage on the myofibrillar protein oxidation and microstructure was the best.

An artificial pathway for trans-4-hydroxy-L-pipecolic acid production from L-lysine in Escherichia coli.

Trans-4-hydroxy-L-pipecolic acid (trans-4-HyPip) is a hydroxylated product of L-pipecolic acid, which is widely used in the pharmaceutical and chemical industries. Here, a trans-4-HyPip biosynthesis module was designed and constructed in Escherichia coli by overexpressing lysine α-oxidase, Δ1-piperideine-2-carboxylase reductase, glucose dehydrogenase, lysine permease, catalase and L-pipecolic acid trans-4-hydroxylase for expanding the lysine catabolism pathway. A total of 4.89 g/L of trans-4-HyPip was generated in shake flasks from 8 g/L of L-pipecolic acid. By this approach, 14.86 g/L of trans-4-HyPip was produced from lysine after 48 h in a 5 L bioreactor. As far as we know, this is the first multi-enzyme cascade catalytic system for the production of trans-4-HyPip using E. coli from L-lysine. Therefore, it can be considered as a potential candidate for the industrial production of trans-4-HyPip in microorganisms.

An Artificial Pathway for N-Hydroxy-Pipecolic Acid Production From L-Lysine in Escherichia coli.

N-hydroxy-pipecolic acid (NHP) is a hydroxylated product of pipecolic acid and an important systemic acquired resistance signal molecule. However, the biosynthesis of NHP does not have a natural metabolic pathway in microorganisms. Here, we designed and constructed a promising artificial pathway in Escherichia coli for the first time to produce NHP from biomass-derived lysine. This biosynthesis route expands the lysine catabolism pathway and employs six enzymes to sequentially convert lysine into NHP. This artificial route involves six functional enzyme coexpression: lysine α-oxidase from Scomber japonicus (RaiP), glucose dehydrogenase from Bacillus subtilis (GDH), Δ1-piperideine-2-carboxylase reductase from Pseudomonas putida (DpkA), lysine permease from E. coli (LysP), flavin-dependent monooxygenase (FMO1), and catalase from E. coli (KatE). Moreover, different FMO1s are used to evaluate the performance of the produce NHP. A titer of 111.06 mg/L of NHP was yielded in shake flasks with minimal medium containing 4 g/L of lysine. By this approach, NHP has so far been produced at final titers reaching 326.42 mg/L by 48 h in a 5-L bioreactor. To the best of our knowledge, this is the first NHP process using E. coli and the first process to directly synthesize NHP by microorganisms. This study lays the foundation for the development and utilization of renewable resources to produce NHP in microorganisms.