In Silico-Predicted Dynamic Oxlipidomics MS/MS Library: High-Throughput Discovery and Characterization of Unknown Oxidized Lipids.

Nontargeted lipidomics using liquid chromatography-tandem mass spectrometry can detect thousands of molecules in biological samples. However, the annotation of unknown oxidized lipids is limited to the structures present in libraries, restricting the analysis and interpretation of experimental data. Here, we describe Doxlipid, a computational tool for oxidized lipid annotation that predicts a dynamic MS/MS library for every experiment. Doxlipid integrates three key simulation algorithms to predict libraries and covers 32 subclasses of oxidized lipids from the three main classes. In the evaluation, Doxlipid achieves very high prediction and characterization performance and outperforms the current oxidized lipid annotation methods. Doxlipid, combined with a molecular network, further annotates unknown chemical analogs in the same reaction or pathway. We demonstrate the broad utility of Doxlipid by analyzing oxidized lipids in ferroptosis hepatocellular carcinoma, tissue samples, and other biological samples, substantially advancing the discovery of biological pathways at the trace oxidized lipid level.

Synergistic Antimicrobial Hybrid Bio-Surface Formed by Self-Assembled BSA Nanoarchitectures with Chitosan Oligosaccharide.

Innovation in green, convenient, and sustainable antimicrobial packaging materials for food is an inevitable trend to address global food waste challenges caused by microbial contamination. In this study, we developed a biogenic, hydrophobic, and antimicrobial protein network coating for food packaging. Experimental results show that disulfide bond breakage can induce the self-assembly of bovine albumin (BSA) into protein networks driven by hydrophobic interactions, and chitosan oligosaccharide (COS) with antimicrobial activity can be stably bound in this network by electrostatic interactions. The inherent antimicrobial activity of COS and the numerous hydrophobic regions on the surface of the BSA-network give the BSA@COS-network significant in vitro antimicrobial ability. More importantly, the BSA@COS-network coating can prolong the onset of spoilage of strawberries in various packaging materials by nearly 3-fold in storage. This study shows how surface functionalization via protein self-assembly is integrated with the biological functioning of natural antibacterial activity for advanced food packaging applications.

Regulation of microbial metabolism on the formation of characteristic flavor and quality formation in the traditional fish sauce during fermentation: a review.

Fish sauce is a special flavored condiment formed by traditional fermentation of low-value fish in coastal areas, which are consumed and produced in many parts of the world, especially in Southeast Asia. In the process of fish sauce fermentation, the diversity of microbial flora and the complex metabolic reactions of microorganisms, especially lipid oxidation, carbohydrate fermentation and protein degradation, are accompanied by the formation of flavor substances. However, the precise reaction of microorganisms during the fersmentation process is difficult to accurately control in modern industrial production, which leads to the loss of traditional characteristic flavors in fermented fish sauces. This paper reviews the manufacturing processes, core microorganisms, metabolic characteristics and flavor formation mechanisms of fermented fish sauces at home and abroad. Various methods have been utilized to analyze and characterize the composition and function of microorganisms. Additionally, the potential safety issues of fermented fish sauces and their health benefits are also reviewed. Furthermore, some future directions and prospects of fermented fish sauces are also reviewed in this paper. By comprehensive understanding of this review, it is expected to address the challenges in the modern production of fish sauce thereby expanding its application in food or diet.

Nanoprecipitates of γ-cyclodextrin/epigallocatechin-3-gallate inclusion complexes as efficient antioxidants for preservation of shrimp surimi products: synthesis, performance and mechanism.

BACKGROUND: Epigallocatechin-3-gallate (EGCG) is well known for excellent chain-breaking antioxidant capability. However, browning by oxidation and aggregation of EGCG is a non-negligible defect that hinders its applications as an antioxidant in various foodstuffs. Therefore, how to eliminate or mitigate browning efficiently, while retaining functionalities as food additive is a challenge in the food industry. RESULTS: Our results demonstrated that EGCG could be anchored within the internal cavity of γ-cyclodextrin (γ-CD) to form an inclusion structure, where hydrophobic interaction, hydrogen bonding, and π-stacking were identified to be the primary drivers. The interplay between two molecules and the steric hindrance from γ-CD could restrict the motion and aggregation of EGCG efficiently, thus alleviating the browning effect. In addition, the conformational adaption of EGCG within the inclusions would result in general decreases in hydrogen-bond dissociation enthalpies for the pyrogallol-type structure on the b ring, thus enhancing the antioxidant capability. In practical application, the nanoscale γ-CD/EGCG inclusion complexes were validated preliminarily as efficient additives in the preservation of shrimp surimi, presenting significant effects on prolonging the shelf-life of products. CONCLUSION: Here, nanoscale γ-CD/EGCG inclusion complexes as alternatives to EGCG were tailored as food antioxidants for the preservation of shrimp surimi products, exerting antioxidant effects while mitigating the browning effects of EGCG on products. Through self-assembly, EGCG would be anchored with the cavity of γ-CD, which could regulate the release modes and restrict the aggregation of EGCG. This facile strategy has great potential in food preservation. © 2023 Society of Chemical Industry.

Supplement of Caulerpa lentillifera polysaccharide by pre-prandial gavage and free feeding demonstrates differences to prevent obesity and gut microbiota disturbance in mice.

BACKGROUND: Caulerpa lentillifera has received extensive attention regarding expansion of its farming and increasing consumption. In our previous study, the structure of C. lentillifera polysaccharide (CLP) was elucidated. However, little information is available about its health effects. In this study, the anti-obesity effect of CLP was investigated by using a high-fat diet-induced obese mice model with two different supplementation methods. RESULTS: In vitro simulated digestion results showed that CLP significantly decreased the lipid digestibility and induced the lipid droplets aggregation in the intestinal stage to inhibit the absorption of lipids. As revealed by 16S ribosomal RNA sequencing and non-targeted metabolomics, supplement of CLP by both pre-prandial gavage and free feeding patterns effectively prevented mice obesity via ameliorating intestinal flora disturbance and regulating bile acids circulation metabolism. Of note was that CLP administration had no effect on short-chain fatty acids production, suggesting the anti-obesity effect was uncorrelated with their production. Moreover, pre-prandial administration of CLP had a better anti-obesity effect in lowering body weight and serum lipid levels, but the free feeding resulted in a higher α-diversity of gut microbiota. CONCLUSION: The findings of this study indicate that CLP could be a potential anti-obesity nutraceutical and that pre-prandial supplement of CLP may be a better intake method to exhibit its hypolipidemic effect. © 2022 Society of Chemical Industry.

Food-grade encapsulated polyphenols: recent advances as novel additives in foodstuffs.

A growing inclination among consumers toward the consumption of natural products has propelled the usage of natural compounds as novel additives. Polyphenols are among the most popular candidates of natural food additives with multiple functionalities and bioactivities but are limited by instability. In this regard, a series of food-grade encapsulated polyphenols has been tailored for incorporating into food formulations as novel additives, which could better satisfy the complicated industry processing. This review seeks to present the most recent discussions regarding their application status in diverse foodstuffs as novel additives, involving functionalities, action mechanisms, and relevant encapsulation technologies. The scientific findings confirm that such novel additives show positive effects on physicochemical, sensory, and nutritional properties as well as the shelf life of diverse food matrices. However, poor heat resistance is still the major defect that restricts their application in thermal processes. Future research should focus on the evaluation of the compatibility and applicability of encapsulated polyphenols in real food processes as well as track and deepen their molecular action mechanisms in the context of complex foodstuffs. Innovation of existing encapsulation technologies should also be concerned in the future to bridge the gap between lab and scale-up production.

Impact of dietary components on enteric infectious disease.

Diets impact host health in multiple ways and an unbalanced diet could contribute to the initiation or progression of a variety of diseases. Although a wealth of information exists on the connections between diet and chronic metabolic diseases such as cardiovascular disease, diabetes mellitus, etc., how diet influences enteric infectious disease still remain underexplored. The review summarizes the current findings on the link between various dietary components and diverse enteric infectious diseases. Dietary ingredients discussed include macronutrients (carbohydrates, lipids, proteins), micronutrients (vitamins, minerals), and other dietary ingredients (phytonutrients and probiotic supplements). We first describe the importance of enteric infectious diseases and the direct and indirect relationship between diet and enteric infectious diseases. Then we discuss the effects of different dietary components on the susceptibility to or progression of enteric infectious disease. Finally, we delineate current knowledge gap and highlighted future research directions. The literature review revealed that different dietary components affect host resistance to enteric infections through a variety of mechanisms. Dietary components may directly inhibit or bind to enteric pathogens, or indirectly influence enteric infections through modulating immune function and gut microbiota. Elucidating the unique repercussions of different diets on enteric infections in this review may help provide dietary guidelines or design dietary interventions to prevent or alleviate enteric infectious diseases.

Comparative metabolomic and transcriptomic analyses revealed the differential accumulation of secondary metabolites during the ripening process of acerola cherry (Malpighia emarginata) fruit.

BACKGROUND: Acerola cherry is a famous functional fruit containing plentiful antioxidants and other nutrients. However, studies on the variations among nutrients during the ripening process of acerola fruit are scare. RESULTS: Comparative metabolomic and transcriptomic analyses were performed and identified 31 331 unigenes and 1896 annotated metabolite features in acerola cherry fruit. K Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that several antioxidant and nutrient-related metabolic pathways, such as the flavonoids, vitamins, carotenoids, amino acids, and fatty acids metabolic pathways, were significantly changed during the ripening process. The metabolites related to the vitamin, carotenoid, and fatty acid metabolic pathways were downregulated during the ripening process. Several flavonoid biosynthesis-related genes (including dihydroflavonol 4-reductase, chalcone synthase, flavanone 3-hydroxylase, and anthocyanidin synthase), were significantly upregulated, suggesting their essential functions in the accumulation of flavonoids in mature fruit. CONCLUSION: Most of the vitamin and carotenoid metabolism-related metabolites significantly accumulated in immature fruit, suggesting that immature acerola fruit is a good material for the extraction of vitamins and carotenoids. For macronutrients, most of the amino acids accumulated in mature fruit and most of the fatty acids greatly accumulated in immature fruit. Our data revealed the differential accumulation of antioxidants and nutrients during the ripening process of acerola cherry fruit. © 2021 Society of Chemical Industry.

A multidrug resistance-associated protein inhibitor is a potential enhancer of the benzyl isothiocyanate-induced apoptosis induction in human colorectal cancer cells.

The increasing drug efflux through the ATP-binding cassette (ABC) transporters is the most plausible mechanism that mediates resistance to the anticancer phytochemicals, such as benzyl isothiocyanate (BITC), as well as chemotherapy drugs. To identify a potential component to overcome this resistance by combinatory utilization, we focused on multidrug resistance-associated proteins (MRPs) pumping various drug metabolites with glutathione as well as the organic anions. The pharmacological treatment of an MRP inhibitor, MK571, significantly potentiated the BITC-induced antiproliferation, coincided with the enhanced accumulation of BITC and glutathione in human colorectal cancer HCT-116 cells. MK571 also enhanced the apoptosis induction as well as activation of the mitogen-activated protein kinases and caspase-3, whereas it did not affect their basal levels. These results suggested that, since MRPs might play a pivotal role in the BITC efflux, MK571 potentiates the BITC-induced antiproliferation in human colorectal cancer cells through inhibition of the glutathione-dependent BITC efflux.

Identification of a xylose-inducible promoter and its application for improving vitamin B12 production in Sinorhizobium meliloti.

Sinorhizobium meliloti 320 is a vitamin B12 (VB12 ) high-producing strain that has been isolated and identified in our previous study. Because the regulatory toolbox for S. meliloti is limited, we searched for new genetic components and identified the two xylose-inducible promoters PA and PB based on a promoter-probe vector with a green fluorescent protein (GFP) as reporter. Compared with the ParaA promoter from S. meliloti, both promoters exhibited higher induced expression and lower basal expression. Subsequently, the influence of glucose or sucrose on the expression of GFP driven by these three promoters was assayed. Glucose repressed all three promoters, and the expression of ParaA was the lowest in the presence of glucose. Although sucrose repressed the expression of PA by 35% and improved the expression of ParaA by 16%, the expression level of PA was the highest and was 13% higher than that of ParaA . Lastly, we overexpressed the hemA gene in the C4 pathway using the PA promoter in S. meliloti 320, and the VB12 production of the engineered strain increased by 11%. The VB12 production was further increased by 11% by adding 0.1% sodium succinate to the culture medium.

Differences in oxidative susceptibilities between glycerophosphocholine and glycerophosphoethanolamine in dried scallop (Argopecten irradians) adductor muscle during storage: an oxidation kinetic assessment.

BACKGROUND: Phospholipids, the main lipid component in marine shellfish, mainly comprise glycerophosphocholine (GPC) and glycerophosphoethanolamine (GPE). GPC and GPE in marine shellfish, especially scallop, carry n-3 long-chain polyunsaturated fatty acids, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), although different types of glycerophospholipids (GP) have different health benefits on human health. Moreover, different GP subclasses such as GPC and GPE have different oxidative susceptibilities in complex food systems. The present study compared the oxidative susceptibilities of GPC and GPE in dried scallop during storage by high-performance liquid chromatography-tandem mass spectrometry and kinetic models, and also investigated the effects of natural phenolic antioxidant on their susceptibilities. RESULTS: The results showed that GPC and GPE molecular species (carrying EPA or DHA) contents in samples continuously reduced during storage at two different temperatures. The first-order kinetic model better reflected the changes of GPC and GPE molecular species (carrying EPA or DHA) in samples than the zero-order kinetic model during storage. According to the oxidation rate (k) obtained from first-order kinetic models, GPE possessed a greater oxidation rate than GPC during storage. Moreover, the results showed that antioxidants of bamboo leaves (AOB, polar polyphenolic antioxidants) significantly decreased the oxidation rates of GPC and GPE molecular species (carrying EPA or DHA) in samples during storage, and GPC could be more effectively protected by AOB compared to GPE. CONCLUSION: The present study provides a practical method for accurately evaluating the oxidative susceptibility of different phospholipid classes in complex food systems. © 2020 Society of Chemical Industry.

Health effects of dietary sulfated polysaccharides from seafoods and their interaction with gut microbiota.

Various dietary sulfated polysaccharides (SPs) have been isolated from seafoods, including edible seaweeds and marine animals, and their health effects such as antiobesity and anti-inflammatory activities have attracted remarkable interest. Sulfate groups have been shown to play important roles in the bioactivities of these polysaccharides. Recent in vitro and in vivo studies have suggested that the biological effects of dietary SPs are associated with the modulation of the gut microbiota. Dietary SPs could regulate the gut microbiota structure and, accordingly, affect the production of bioactive microbial metabolites. Because of their differential chemical structures, dietary SPs may specifically affect the growth of certain gut microbiota and associated metabolite production, which may contribute to variable health effects. This review summarizes the latest findings on the types and structural characteristics of SPs, the effects of different processing techniques on the structural characteristics and health effects of SPs, and the current understanding of the role of gut microbiota in the health effects of SPs. These findings might help in better understanding the mechanism of the health effects of SPs and provide a scientific basis for their application as functional food.

Sensory evaluation of fresh/frozen mackerel products: A review.

Mackerel has received considerable attention in the global food market as one of the most important pelagic commercial fish species. The quality of mackerel is influenced by species, season, fishing area, nutritional status, catching method, handling, and storage. Due to the mackerel's perishability, its quality is mainly measured by sensory procedures. Although considerable effort has been made to explore quick and reliable quality analysis, developing a practical and scientific sensory evaluation of mackerel has been an active ongoing study area to meet the quality evaluation demand of the industry. Different sensory evaluation methods have been used to assess the mackerel fish quality, including Palatability and Spoilage test, Torry scheme, EU scheme, Quality Index Method, Catch damage index and Processed fish damage index, Affective test, Discriminative test, and Descriptive test. Each method has its strength and weakness. Despite mackerel sensory evaluation protocols having undergone partial harmonization, specific sample process needs to be carefully followed to minimize the change during sample preparation. This review summarizes the sensory evaluation methods in mackerel research, the factors affecting sensory evaluation, and then updates the latest advances in mackerel sensory evaluation and offers guidance for presenting its application in the mackerel chain. Also, each technique's advantages and limitations are discussed. In our opinion, the future trends for sensory evaluation of mackerel should be consumer-centric.

Improving oxidative stability and release behavior of docosahexaenoic acid algae oil by microencapsulation.

BACKGROUND: Spray-dried docosahexaenoic acid algae oil (DHA AO) microcapsules were prepared using whey protein isolate and glucose syrup (WPI + GS), or sodium starch octenylsuccinate and glucose syrup (SSOS + GS), or whey protein isolate and lactose (WPI + L). The effect of the formulations on encapsulation properties, oxidative protection and in vitro oil release pattern of the resulting microencapsulates was investigated. RESULTS: A high encapsulation efficiency of over 98% of DHA AO was obtained for microcapsules with all three wall materials. Among the wall materials, SSOS + GS exhibited a better micro-particulation ability reflected by more uniform size and smoother surface of the formed microcapsules and no agglomerates. DHA AO microcapsules with all the wall materials showed good protection of the oil from oxidation during storage with an increasing order of WPI + GS, SSOS + GS and WPI + L. Moreover, microencapsulation significantly increased the release of DHA AO in the intestinal phase of the in vitro digestion process with an increasing order of SSOS + GS, WPI + GS and WPI + L, indicating the increased stability of the oil in the highly acidic gastric environment and the enhanced lipid digestibility in the small intestine. CONCLUSIONS: The results suggest that it is possible to transform a highly oxidizable liquid functional food ingredient such as DHA AO into a stable and easy-to-handle solid powder through spray drying with properly selected wall materials. © 2020 Society of Chemical Industry.

Zinc-Chelating Mechanism of Sea Cucumber (Stichopus japonicus)-Derived Synthetic Peptides.

In this study, three synthetic zinc-chelating peptides (ZCPs) derived from sea cucumber hydrolysates with limited or none of the common metal-chelating amino-acid residues were analyzed by flame atomic absorption spectroscopy, circular dichroism spectroscopy, size exclusion chromatography, zeta-potential, Fourier transform infrared spectroscopy, Raman spectroscopy and nuclear magnetic resonance spectroscopy. The amount of zinc bound to the ZCPs reached maximum values with ZCP:zinc at 1:1, and it was not further increased by additional zinc presence. The secondary structures of ZCPs were slightly altered, whereas no formation of multimers was observed. Furthermore, zinc increased the zeta-potential value by neutralizing the negatively charged residues. Only free carboxyl in C-terminus of ZCPs was identified as the primary binding site of zinc. These results provide the theoretical foundation to understand the mechanism of zinc chelation by peptides.

Antioxidative Peptides from Proteolytic Hydrolysates of False Abalone (Volutharpa ampullacea perryi): Characterization, Identification, and Molecular Docking.

Antioxidative peptides were produced from false abalone (Volutharpa ampullacea perryi) using enzymatic hydrolysis. Trypsin produced the most bioactive hydrolysates with the highest scavenging ABTS+• free radicals compared to pepsin, alcalase, neutrase, and flavourzyme. The response surface methodology studies on trypsin hydrolysis indicated that the hydrolysis temperature, time, and pH were interacted with each other (p < 0.05), and the optimal conditions were hydrolysis at 51.8 °C for 4.1 h, pH 7.7 and the maximum predicted hydrolysis degree was 13.18% and ABTS+• scavenging activity of 79.42%. The optimized hydrolysate was subjected to ultrafiltration fractionation, and the fraction with MW < 3 kDa showed the highest ABTS+• scavenging activity. There were 193 peptide sequences identified from this peptide fraction and 133 of them were successfully docked onto human myeloperoxidase (MPO), an enzyme involved in forming reactive oxidants in vivo. The highest scored peptide, no. 39, consists of DTETGVPT. Its structure and molecular interactions with MPO active site were compared with previously characterized peptide hLF1-11. The interactions between peptide no. 39 and MPO include electrostatic charge, hydrogen bonds, and covalent bonds. The antioxidative peptide produced in this research may exert antioxidant activity in vivo due to its potential inhibition effect on MPO.

Quantitative Proteome Reveals Variation in the Condition Factor of Sea Urchin Strongylocentrotus nudus during the Fishing Season Using an iTRAQ-based Approach.

To investigate the variation in the condition factor of the sea urchin Strongylocentrotus nudus (S. nudus), gonads were collected in May (MAY), June (JUN), and July (JUL), at the beginning (AUG-b) and end of August (AUG-e). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) detection of the gonads revealed an obvious enhancement of the band at about 37 kDa from July, which was identified as transforming growth factor-beta-induced protein ig-h3 (TGFBI) by nanoLC-ESI-MS/MS. Gonadal proteins were identified by isobaric tagging for relative and absolute quantitation (iTRAQ), and regulation of the identified proteins in pairs of the collected groups was observed. A total of 174 differentially expressed proteins (DEPs) were identified. Seven of the DEPs showed significant correlations with both the gonad index (GI) and protein content. These correlations included 6-phosphogluconate dehydrogenase, decarboxylating isoform X2 (6PGD), CAD protein, myoferlin isoform X8, ribosomal protein L36 (RL36), isocitrate dehydrogenase [NADP], mitochondrial isoform X2 (IDH), multifunctional protein ADE2 isoform X3, sperm-activating peptides (SAPs) and aldehyde dehydrogenase, and mitochondrial (ALDH). However, TGFBI had no correlation with gonad index (GI) or protein content. 6PGD, IDH, multifunctional protein ADE2 isoform X3, and ALDH were shown to interact with each other and might play key roles in changing the condition factor of S. nudus gonads.

RNA Sequencing Analysis to Capture the Transcriptome Landscape during Tenderization in Sea Cucumber Apostichopus japonicus.

Sea cucumber (Apostichopus japonicus) is an economically significant species in China having great commercial value. It is challenging to maintain the textural properties during thermal processing due to the distinctive physiochemical structure of the A. japonicus body wall (AJBW). In this study, the gene expression profiles associated with tenderization in AJBW were determined at 0 h (CON), 1 h (T_1h), and 3 h (T_3h) after treatment at 37 °C using Illumina HiSeq™ 4000 platform. Seven-hundred-and-twenty-one and 806 differentially expressed genes (DEGs) were identified in comparisons of T_1h vs. CON and T_3h vs. CON, respectively. Among these DEGs, we found that two endogenous proteases-72 kDa type IV collagenase and matrix metalloproteinase 16 precursor-were significantly upregulated that could directly affect the tenderness of AJBW. In addition, 92 genes controlled four types of physiological and biochemical processes such as oxidative stress response (3), immune system process (55), apoptosis (4), and reorganization of the cytoskeleton and extracellular matrix (30). Further, the RT-qPCR results confirmed the accuracy of RNA-sequencing analysis. Our results showed the dynamic changes in global gene expression during tenderization and provided a series of candidate genes that contributed to tenderization in AJBW. This can help further studies on the genetics/molecular mechanisms associated with tenderization.

Isolation and identification of zinc-chelating peptides from sea cucumber (Stichopus japonicus) protein hydrolysate.

BACKGROUND: Zinc is known to play an essential role in the biological activities in the human body. In this study, a zinc-chelating peptide (ZCP) produced by Alcalase-assisted hydrolysis of the body wall of sea cucumber was isolated and identified. The ZCP was purified stepwise by ultrafiltration, anion-exchange chromatography, and gel filtration chromatography, in conjunction with ultraviolet-visual (UV-visual) spectrophotometry, which was used to analyze each purified fraction. RESULTS: Analysis of the purified ZCP revealed that its zinc-chelating ability was 33.31%. Analysis of isothermal titration calorimetry suggested that the binding of ZCP and zinc (N ≈ 2) was endothermic, with weak binding affinity. Fourier transform infrared spectroscopy spectra (FTIR) indicated that carboxylic and amide groups in ZCP were the primary binding sites of Zn. Sequencing the result by ultra-performance liquid chromatography-quadrupole/time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) showed that a representative ZCP had the sequence WLTPTYPE with a molecular weight of 1005.5 Da. CONCLUSION: These results provide a promising foundation for the production of zinc supplements from sea-cucumber-derived ZCPs. © 2019 Society of Chemical Industry.

Genetic engineering of Escherichia coli to improve L-phenylalanine production.

BACKGROUND: L-phenylalanine (L-Phe) is an essential amino acid for mammals and applications expand into human health and nutritional products. In this study, a system level engineering was conducted to enhance L-Phe biosynthesis in Escherichia coli. RESULTS: We inactivated the PTS system and recruited glucose uptake via combinatorial modulation of galP and glk to increase PEP supply in the Xllp01 strain. In addition, the HTH domain of the transcription factor TyrR was engineered to decrease the repression on the transcriptional levels of L-Phe pathway enzymes. Finally, proteomics analysis demonstrated the third step of the SHIK pathway (catalyzed via AroD) as the rate-limiting step for L-Phe production. After optimization of the aroD promoter strength, the titer of L-Phe increased by 13.3%. Analysis of the transcriptional level of genes involved in the central metabolic pathways and L-Phe biosynthesis via RT-PCR showed that the recombinant L-Phe producer exhibited a great capability in the glucose utilization and precursor (PEP and E4P) generation. Via systems level engineering, the L-Phe titer of Xllp21 strain reached 72.9 g/L in a 5 L fermenter under the non-optimized fermentation conditions, which was 1.62-times that of the original strain Xllp01. CONCLUSION: The metabolic engineering strategy reported here can be broadly employed for developing genetically defined organisms for the efficient production of other aromatic amino acids and derived compounds.