Production Of Free Amino Acids Research Articles

Astral-based DIA proteomics explored the flavor enhancement mechanism of Chinese traditional smoked bacon by staphylococcal co-fermentation

The proteolysis pattern during mixed fermentation of Staphylococcus cohnii WX-M8 and S. saprophyticus MY-A10 on Chinese bacon was still unknown. In this study, the changing laws of protein degradation products during staphylococcal mixed fermentation were analyzed, followed by an investigation of endogenous enzymes and cellular components, and finally an examination of flavor profiles. Results indicated that mixed fermentation improved protein degradation and promoted the production of peptides and free amino acids (FAAs). Proteolysis of S. saprophyticus MY-A10 was non-specific, and it promoted protein degradation by cooperating with cathepsin L1. S. cohnii WX-M8 was specific and acted mainly with calpain-3 in the thin filament. The fulfillment of S. cohnii WX-M8 function was enhanced in the presence of S. saprophyticus MY-A10. Mixed fermentation showed synergism with endogenous peptidases in degrading peptides to small-molecule peptides or FAAs and complementarity with endogenous dehydrogenases in converting FAAs to volatile organic compounds (VOCs).

Research on microbial diversity and nutritional flavor formation of Xianju wheat paste

AbstractBACKGROUNDXianju wheat paste, a traditional condiment in Hubei Province, China, possesses nutritional value and a distinctive taste profile. Nevertheless, there remains a dearth of comprehensive comprehension regarding the intricate interplay between the microbial population and its nutritional profile in Xianju wheat paste.RESULTSIt was determined that Xianju wheat paste harbors predominant microbial genera such as Bacillus, Staphylococcus and Aspergillus. The findings from high‐throughput sequencing analysis revealed the presence of 11 bacterial genera in Xianju wheat paste, with relative abundances exceeding 1%. These genera included Bacillus, Staphylococcus, Brevibacterium, Lactobacillus, Allorhizobium‐Neorhizobium‐Pararhizobium‐Rhizobium, Enterobacteriaceae, Pantoea, Brachybacterium, Klebsiella, Escherichia‐Shigella and Ochrobactrum. Furthermore, six fungal genera were identified with relative abundances greater than 1%, specifically Aspergillus, Zygosaccharomyces, Cutaneotrichosporon, Candida, Millerozyma and Rhizopus. The correlation analysis indicated a significant impact of the microbial community and nutritional flavor on the second phase of fermentation in Xianju wheat paste. The predominant bacterium Bacillus in Xianju wheat paste facilitated the production of free amino acids, whereas Staphylococcus exhibited a negative correlation with free amino acid levels. The quantity of volatile compounds in Xianju wheat paste progressively increased during fermentation, with the presence of typical aroma compounds 4‐vinyl‐2‐methoxyphenol significantly associated with Bacillus. This indicated Bacillus notably enhanced the production of aromatic substances. Furthermore, a strong correlation was observed between Staphylococcus and the 18 volatile organic compounds, highlighting their substantial contribution to these aroma compounds.CONCLUSIONThis research indicates that the presence of microbial communities significantly contributes to the development of the nutritional flavor profile in Xianju wheat paste. © 2024 Society of Chemical Industry.

Hemp Seed Protein Hydrolysate Enriched with γ-Aminobutyric Acid and Peptides by Microbial Bioconversion

Hemp seed protein (HSP), a by-product of hemp oil processing, was converted into a functional protein ingredient enriched with γ-aminobutyric acid (GABA) and peptides through a two-step microbial fermentation process. To enhance peptide and free amino acid production from HSP, it was hydrolyzed using alkaline protease produced by Bacillus subtilis HA. The HSP was hydrolyzed at a degree of 40% at 55 °C for 24 h, yielding a pH of 6.55, an acidity of 1.22%, and 205.45 mg% tyrosine equivalents. This process resulted in the production of low molecular-weight peptides. (<5000 Da) The total amino acid content and branched-chain amino acids (leucine, isoleucine, and valine) were 6.78 mg/g and 1.47 mg/g. Subsequently, the production of γ-aminobutyric acid (GABA) in the HSP hydrolysate was optimized through co-fermentation with lactic acid bacteria in the presence of 5% MSG at 30 °C for 5 days. The serial co-fermented HSP hydrolysate exhibited a GABA content of 33.98 mg/g and a viable bacterial count of 9.51 log CFU/mL for Lb. plantarum KS2020. This serial co-fermentation process, combining proteolysis and lactic acid fermentation, not only increased the peptide content but also promoted GABA accumulation, positioning HSP hydrolysate as a promising candidate for functional foods with potential health benefits.

Transcriptomic and Metabolomic Analyses of Soybean Protein Isolate on Monascus Pigments and Monacolin K Production.

Monascus pigments (MPs) and monacolin K (MK) are important secondary metabolites produced by Monascus spp. This study aimed to investigate the effect of soybean protein isolate (SPI) on the biosynthesis of MPs and MK based on the analysis of physiological indicators, transcriptomes, and metabolomes. The results indicated that the growth, yellow MPs, and MK production of Monascus pilosus MS-1 were significantly enhanced by SPI, which were 8.20, 8.01, and 1.91 times higher than that of the control, respectively. The utilization of a nitrogen source, protease activity, the production and utilization of soluble protein, polypeptides, and free amino acids were also promoted by SPI. The transcriptomic analysis revealed that the genes mokA, mokB, mokC, mokD, mokE, mokI, and mokH which are involved in MK biosynthesis were significantly up-regulated by SPI. Moreover, the glycolysis/gluconeogenesis, pyruvate metabolism, fatty acid degradation, tricarboxylic acid (TCA) cycle, and amino acid metabolism were effectively up-regulated by SPI. The metabolomic analysis indicated that metabolisms of amino acid, lipid, pyruvate, TCA cycle, glycolysis/gluconeogenesis, starch and sucrose, and pentose phosphate pathway were significantly disturbed by SPI. Thus, MPs and MK production promoted by SPI were mainly attributed to the increased biomass, up-regulated gene expression level, and more precursors and energies.

Effects of ultrasonic-assisted marinating on degradation of beef protein and formation of flavor precursors

In the process of meat production, marinating has a great influence on the flavor of the final product. In this study, the effect of ultrasound-assisted dry-marinating on protein degradation, structural changes, and formation of flavor precursor substances were investigated for 12 h and 24 h. The increase in protein degradation index and protease activities under ultrasound promoted the production of free amino acids. After sonication, the microstructure of muscle tissue was loose, and the relative content of β-turn and β-sheet were reduced, indicating that ultrasonic treatment facilitated the oxidation and hydrolysis of proteins. The increased proteolysis could be reflected in increased degradation of myosin light chains and troponin-T. It’s worth noting that ultrasonic-assisted dry-marinating 90 min and continued static marinating until 12 h could achieve the treatment of static marinating for 24 h. These results indicated that ultrasound-assisted dry-marinating could effectively improve the marinating efficiency by promoting protein degradation, affecting the formation of flavor precursors in meat.

New insights into the dominance of mixed fermentation of Staphylococcus cohnii and Staphylococcus saprophyticus in Chinese bacon: Complete genomic and comparative genomic perspectives

Previous studies have demonstrated that Staphylococcus cohnii WX_M8 and S. saprophyticus MY_A10 significantly enhanced the flavor of Chinese bacon in a mixed fermentation. However, due to the complexity of the processing, the contribution of the bacteria is deceptive when investigating only the phenotypic changes at the time of fermentation. In order to clarify the metabolic mechanisms of mixed fermentation, a technological characterization, whole genome and comparative genomics analysis, and metabolites were approached in this study. Results showed that differences in tolerance characteristics existed between WX_M8 and MY_A10. And the genomes of both the two strains consisted of one chromosome and four circular plasmids. Their genome sizes were 2.74 Mp and 2.62 Mp, the GC contents were 32.45% and 33.18%, and the predicted coding genes (CDS) were 2564 and 2541, respectively. Based on the annotation of gene functions and assessment of metabolic pathways in the KEGG database, WX_M8 and MY_A10 strains were found to harbor complete protein degradation and amino acid metabolic pathways, pyruvate and butanol metabolic pathways, and isoleucine metabolic pathways, and their diverse enzyme-encoding genes superimposed the metabolic functions, whereas the alcohol dehydrogenase genes, adh and frmA, achieved complementary functions in the production of esters. Comparative genomics analysis revealed a diversity of encoding genes of aminotransferases and a greater metabolism for sulfur-containing amino acids, aromatic amino acids, and branched-chain amino acids in the mixed fermentation of strains WX_M8 and MY_A10. Metabolites analysis showed that MY_A10 focused on the production of soluble peptides and free amino acids (FAAs), while WX_M8 focused on volatile organic compounds (VOCs), resulting in a significant enhancement of the flavor of Chinese bacon when the two were mixed fermented. This result may provide direction for strains WX_M8 and MY_A10 to be used as starter cultures and targeted to regulate flavor.

Interspecific Differences in Carbon and Nitrogen Metabolism and Leaf Epiphytic Bacteria among Three Submerged Macrophytes in Response to Elevated Ammonia Nitrogen Concentrations.

Submerged macrophytes in eutrophic aquatic environments adapt to changes in ammonia nitrogen (NH4-N) levels by modifying their levels of free amino acids (FAAs) and soluble carbohydrates (SCs). As symbionts of submerged macrophytes, epiphytic bacteria have obvious host specificity. In the present study, the interspecific differences in the FAA and SC contents of Hydrilla verticillata (Linn. f.) Roylep, Vallisneria natans Hara and Chara braunii Gmelin and their leaf epiphytic bacterial communities were assessed in response to increased NH4-N concentrations. The results revealed that the response of the three submerged macrophytes to NH4-N stress involved the consumption of SCs and the production of FAAs. The NH4-N concentration had a greater impact on the variation in the FAA content, whereas the variation in the SC content was primarily influenced by the species. At the phylum level, the relative abundance of Nitrospirota on the leaves exhibited specific differences, with the order H. verticillata > V. natans > C. braunii. The dominant genera of epiphytic bacteria with denitrification effects on V. natans, H. verticillata and C. braunii leaves were Halomonas, Acinetobacter and Bacillus, respectively. When faced with NH4-N stress, the variation in epiphytic bacterial populations associated with ammonia oxidation and denitrification among submerged macrophytes could contribute to their divergent responses to heightened nitrogen levels.

Characterization and Hydrolysis Mechanism Analysis of a Cold-Adapted Trypsin-Like Protease from Antarctic Krill.

Cold-adapted proteases are capable of efficient protein hydrolysis at reduced temperatures, which offer significant potential applications in the area of low temperature food processing. In this paper, we attempted to characterize cold-adapted proteases from Antarctic krill. Antarctic krill possesses an extremely active autolytic enzyme system in their bodies, and the production of peptides and free amino acids accompanies the rapid breakdown of muscle proteins following the death. The crucial role of trypsin in this process is recognized. A cold-adapted trypsin named OUC-Pp-20 from Antarctic krill genome was cloned and expressed in Pichia pastoris. Recombinant trypsin is a monomeric protein of 26.8 ± 1.0 kDa with optimum reaction temperature at 25 °C. In addition, the catalytic specificity of OUC-Pp-20 was assessed by identifying its hydrolysis sites through LC-MS/MS. OUC-Pp-20 appeared to prefer Gln and Asn at the P1 position, which is an amino acid with an amide group in its side chain. Hydrolysis reactions on milk and shrimp meat revealed that it can effectively degrade allergenic components in milk and arginine kinase in shrimp meat. These findings update the current knowledge of cold-adapted trypsin and demonstrate the potential application of OUC-Pp-20 in low temperature food processing.

Recent innovations in fertilization with treated digestate from food waste to recover nutrients for arid agricultural fields

This study aims to explore the development of sustainable fertilizers from waste materials of a biogas plant and a brewery. These wastes, rich in organic carbon and nitrogen, were processed with sulfuric(VI) and phosphoric(V) acid mixture, facilitating the production of free amino acids and achieving waste sanitization. This treatment produced by-products, which extended the range of possible applications. The highest concentration of free amino acids (360 mg/l) was achieved through hydrolyzing with a 40% concentration medium over 24 h. In this case, the maximum levels were recorded for beta-alanine (69.3 mg/l), glycine (46.8 mg/l), isoleucine (43.5 mg/l), proline (36.2 mg/l), and valine (31.5 mg/l). The study presents two fertilizer technologies, with and without micronutrients, that satisfy European Parliament Regulation 2019/1009 (Ntot > 2%, Norg > 0.5%, Corg > 3%). Bioavailability of nutrients in the formulations ranged from 60 to 100%. The efficacies of these fertilizers were evaluated in 30-day pot trials with various plant species, with both single application and fertigation tested. Multielement analysis confirmed high nutrient transfer in the soil-plant system, and the inclusion of micronutrients led to biofortification of plant biomass in Cu (48.3 ± 7.2 mg/kg), Mn (249 ± 37 mg/kg), Zn (164 ± 25 mg/kg), and Fe (211 ± 32 mg/kg). These sustainable fertilizers present an alternative to traditional, non-renewable fertilizers and offer promising solutions for precision agriculture and environmentally conscious production.Graphical abstract

Investigating the effectiveness of coacervates produced from conjugated and unconjugated Spirulina protein in delivering unstable oil to the intestinal phase of digestion

This study investigated the potential of complex coacervates produced using Spirulina protein concentrate (SPC) conjugated with maltodextrin (MD) and carrageenan (CG) for encapsulating and delivering sensitive oils. A wet-heating Maillard reaction was employed to conjugate SPC with MD, followed by coacervation with CG to form the conjugate-based coacervates. Additionally, a mixture of unconjugated SPC and MD was coacervated with CG to produce mixture-based coacervates. Both types of coacervates were utilised as wall materials for encapsulating canola oil. The in-vitro digestion of the resulting microcapsules was assessed in oral, gastric, and intestinal phases, focusing on physicochemical parameters such as droplet size, zeta-potential, microstructure, proteolysis, oil release and lipolysis. The findings revealed that microcapsules prepared using both (SPC-MD mixture)-CG and (SPC-MD conjugate)-CG coacervates were remarkably stable against gastric digestion, as evidenced by the minimal production of free amino acids (15 mM). Most of the encapsulated oil (62–67%) was released during the intestinal phase due to the breakdown of the coacervates. Notably, the microcapsules produced with (SPC-MD conjugate)-CG coacervates demonstrated a lower degree of lipolysis (41.77% free fatty acid content) compared to those prepared with (SPC-MD mixture)-CG coacervates (53.35% free fatty acid content). These results highlight the potential of complex coacervates produced using conjugated SPC as promising materials for the encapsulation and delivery of sensitive oils.

Effects of Co-Fermentation of Lactiplantibacillus plantarum and Saccharomyces cerevisiae on Digestive and Quality Properties of Steamed Bread.

The leavening of wheat-based steamed bread is carried out either with a pure yeast culture or with traditional starter cultures containing both lactic acid bacteria and yeast/mold. The use of variable starter cultures significantly affects steamed bread's quality attributes, including nutritional profile. In this paper, differences in physicochemical properties, the type of digested starch, the production of free amino acids, and the specific volume of steamed bread under three fermentation methods (blank, yeast, and LP-GM4-yeast) were compared. The digestion characteristics (protein and starch hydrolysis) of steamed bread produced by using either yeast alone or a combination of Lactiplantibacillus plantrum and yeast (LP-GM4-yeast) were analyzed by an in vitro simulated digestion technique. It was found that the specific volume of steamed bread fermented by LP-GM4-yeast co-culture was increased by about 32%, the proportion of resistant starch was significantly increased (more than double), and soluble protein with molecular weight of 30-40 kDa was significantly increased. The results of this study showed that steamed bread produced by LP-GM4-yeast co-culture is more beneficial to human health than that by single culture.

Metabolomics and metatranscriptomics reveal the influence mechanism of endogenous microbe (Staphylococcus succinus) inoculation on the flavor of fermented chili pepper

This study integrated metabolomic and metatranscriptomic techniques to examine how the endogenous microbe, Staphylococcus succinus, influenced the essential flavor of fermented chili peppers. The mechanisms governing spontaneous fermentation and S. succinus-inoculated fermentation were also elucidated. Esters (e.g., ethyl undecanoate, isoamyl acetate, and methyl salicylate), terpenes (e.g., terpinen-4-ol), and alcohols (e.g., α-terpineol, linalool, and 4-methyl-3-heptanol) were found to be the key aroma-active compounds, aspartic acid (Asp) and glutamic acid (Glu) were identified as primary flavoring free amino acids. Notably, during the early stages of S. succinus-inoculated fermentation, the production of these essential metabolites was abundant, while their gradual increase over time was observed in the case of spontaneous fermentation. Metatranscriptomic analysis revealed that S. succinus inoculation could up-regulate genes related to glycolysis, amino acid metabolism, and aroma compound synthesis. These changes sequentially boosted the production of sweet and umami free amino acids, enhanced organic acid levels, increased unique aroma compound generation, and further improved the flavor and quality of the fermented chili peppers. Therefore, S. succinus inoculation can augment the sensory quality of fermented chili peppers, making this strain a promising candidate for Sichuan pickle fermentation starters.

Copper toxicity leads to accumulation of free amino acids and polyphenols in Phaeodactylum tricornutum diatoms

This work is focused on the effect of lethal and sub-lethal copper (Cu) concentrations on the free amino acid and polyphenol production by the marine diatom Phaeodactylum tricornutum (P. tricornutum) after 12, 18, and 21 days of exposure. The concentrations of 10 amino acids (arginine, aspartic acid, glutamic acid, histidine, lysine, methionine, proline, valine, isoleucine, and phenylalanine) and 10 polyphenols (gallic acid, protocatechuic acid, p-coumaric acid, ferulic acid, catechin, vanillic acid, epicatechin syringic acid, rutin, and gentisic acid) were measured by RP-HPLC. Under lethal doses of Cu, free amino acids reached levels significantly higher than those in the control cells (up to 21.9 times higher), where histidine and methionine showed the highest increases (up to 37.4 and 65.8 times higher, respectively). The total phenolic content also increased up to 11.3 and 5.59 times higher compared to the reference cells, showing gallic acid the highest increase (45.8 times greater). The antioxidant activities of cells exposed to Cu were also enhanced with increasing doses of Cu(II). They were evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging ability (RSA), cupric ion reducing antioxidant capacity (CUPRAC), and ferric reducing antioxidant power (FRAP) assays. Malonaldehyde (MDA) exhibited the same tendency: cells grown at the highest lethal Cu concentration yielded the highest MDA level. These findings reflect the involvement of amino acids and polyphenols in protective mechanisms to overcome the toxicity of copper in marine microalgae.Graphical

Conversion of anaerobic digestates from biogas plants: Laboratory fertilizer formulation, scale-up and demonstration of applicative properties on plants

This research paper presents a zero-waste method for preparing a fertilizer based on digestate from food residues. The waste was treated with a mixture of inorganic acids (sulfuric acid and phosphoric acid). The process allowed for sanitization with simultaneous production of free amino acids and short peptides. In preliminary studies, the effect of the order of addition of components (digestate hydrolysate, ash from incineration of sewage sludge) on nitrogen loss was observed during the neutralization/granulation process. The slow addition of ash to the hydrolysate minimizes nitrogen loss (0.6 g N/kg fertilizer) six times compared to the reverse order. Based on the optimal conditions, a large-scale fertilizer was produced, meeting the quality requirements of the EU Regulation for solid inorganic macronutrient fertilizer - PFC 1(C)(I)(a)(i). The physicochemical properties of the final fertilizer were investigated: microbiological analysis, granule strength tests, and bioavailability and leachability of nutrients in extraction tests. The effectiveness of the fertilizer was confirmed in 30-day pot tests, with results similar to those of the mineral fertilizer. The fertilizer obtained can be a substitute for traditional mineral fertilizers, which is a promising solution in the situation of rising natural gas prices and the availability of fossil raw materials.

Effects of Brines and Containers on Flavor Production of Chinese Pickled Chili Pepper (Capsicum frutescens L.) during Natural Fermentation.

The effects of (fresh/aged) brine and (pool/jar) containers on the flavor characteristics of pickled chili peppers were investigated based on a multivariate analysis integrated with kinetics modeling. The results showed that the effect of brine on organic acid, sugar, and aroma was more dominant than that of containers, while free amino acids production was more affected by containers than brines. Chili pepper fermented using aged brine exhibited higher acidity (3.71−3.92) and sugar (7.92−8.51 mg/g) than that using fresh brine (respective 3.79−3.96; 6.50−9.25 mg/g). Besides, chili peppers fermented using pool containers showed higher free amino acids content (424.74−478.82 mg/100 g) than using a jar (128.77−242.90 mg/100 g), particularly with aged brine. As for aroma, the number of volatiles in aged brine was higher (88−96) than that in fresh brine (76−80). The contents of the esters, alcohols, and ketones were significantly higher in the aged brine samples than those in fresh brine (p < 0.05), while terpenes in chili pepper fermented using the pool were higher than those using the jar. In general, jar fermentation with aged brine contributed more flavor to pickled chili peppers than other procedures.

Overexpression of the FERONIA receptor kinase MdMRLK2 confers apple drought tolerance by regulating energy metabolism and free amino acids production.

Drought is a major abiotic stress limiting the growth and production of apple trees worldwide. The receptor-like kinase FERONIA is involved in plant growth, development and stress responses; however, the function of FERONIA in apple under drought stress remains unclear. Here, the FERONIA receptor kinase gene MdMRLK2 from apple (Malus domestica) was shown to encode a plasma membrane-localized transmembrane protein and was significantly induced by abscisic acid and drought treatments. 35S::MdMRLK2 apple plants showed less photosystem damage and higher photosynthetic rates compared with wild-type (WT) plants, after withholding water for 7days. 35S::MdMRLK2 apple plants also had enhanced energy levels, activated caspase activity and more free amino acids, than the WT, under drought conditions. By performing yeast two-hybrid screening, glyceraldehyde-3-phosphate dehydrogenase and MdCYS4, a member of cystatin, were identified as MdMRLK2 interaction partners. Moreover, under drought conditions, the 35S::MdMRLK2 apple plants were characterized by higher abscisic acid (ABA) content. Overall, these findings demonstrated that MdMRLK2 regulates apple drought tolerance, probably via regulating levels of energetic matters, free amino acids and ABA.

Kinetic study of some flavor and bioactive compounds during fermentation of Parkia biglobosa

Seeds of Parkia biglobosa, also called African locust bean seeds (ALBSs), are used to produce a condiment by subjecting them to cooking, alkaline fermentation, and drying. The objective of this work was to monitor the evolution of the composition of ALBS during spontaneous fermentation. Results showed that the seeds at initial time contained mainly proteins (45%) and lipids (36%). During fermentation, the conversion of this initial composition took place to produce ammonium, free amino acids, and short-chain fatty acids (SCFAs) that accounted for 2%, 10%, and 1.5% dry basis, respectively, which explains the typical flavor of fermented ALBS. This release was also concomitant with biogenic amine production. Along with ammonium and SCFA release, pyrazines were the major volatile organic compounds. Composition modulation was well represented by a Verhulst equation in which the dynamic could be modified by the initial composition of the seed, the pre-cooking step, and the fermentation conditions (temperature and aeration). Practical applications For nutritional and sustainability reasons, re-integration of vegetable in general and legumes, in particular in our diet, is a current trend. However, only dish that are appealing to consumers are concretely adopted. Fermentation of legumes is interesting in more than one way. It does not involve heat, so thermolabile molecules can be preserved and the financial and energetic costs are low. Biological interesting molecules are produced from the metabolism of microorganisms, and among them, volatile organic compounds that contribute to enhance the flavor of legume dishes. Soumbala is a meal and/or condiment obtained from the fermentation of African locust bean (Parkia biglobosa) seeds (ALBSs), a legume widely spread in west Africa. The final typical sensory properties are known to be due to the alkaline fermentation step by Bacillus species. This paper presents the kinetics of release of various compounds during spontaneous fermentation that may impact the sensory and nutritional properties of the final product. More precisely, the dynamic of the production of free amino acids, free fatty acids, volatile compounds, and biogenic amines are presented and discussed to give insights into the succession of important reactions to control the sensory and nutritional qualities of fermented ALBS (FALBS). This paper shows that further research are necessary to better understand and control the quality of not only FALBS but also similar protein-rich legumes to modulate the quality of the final products.

Effect of starter cultures and spices on physicochemical properties and microbial communities of fermented fish (Suanyu) after fermentation and storage

Suanyu is one of the traditional Chinese fermented fish products with a pleasant flavor. The present study sought to evaluate the effects of starter cultures and spices on the physicochemical properties and microbial flora of Suanyu after fermentation and storage. The results indicated that fermentation with mixed starter cultures could lower the pH and inhibit TVB-N formation in Suanyu after fermentation and during storage. Moreover, inoculation of starter cultures could promote free amino acids production during storage. Additionally, inoculation of mixed starter cultures and spices mixture could effectively inhibit the formation of most biogenic amines in Suanyu, revealing the significant role of mixed starter cultures. Staphylococcus, Macrococcus, and Lactobacillus were the core microbial genus in Suanyu. The inoculation of starter cultures could inhibit Enteroccoccus and other microorganisms, while spices introduced Bacillus into Suanyu. The study results provide a theoretical basis for the industrial production and storage of Suanyu.

Calpain-mediated proteolytic production of free amino acids in vascular endothelial cells augments obesity-induced hepatic steatosis

Free amino acids that accumulate in the plasma of patients with diabetes and obesity influence lipid metabolism and protein synthesis in the liver. The stress-inducible intracellular protease calpain proteolyzes various substrates in vascular endothelial cells (ECs), although its contribution to the supply of free amino acids in the liver microenvironment remains enigmatic. In the present study, we showed that calpains are associated with free amino acid production in cultured ECs. Furthermore, conditioned media derived from calpain-activated ECs facilitated the phosphorylation of ribosomal protein S6 kinase (S6K) and de novo lipogenesis in hepatocytes, which were abolished by the amino acid transporter inhibitor, JPH203, and the mammalian target of rapamycin complex 1 inhibitor, rapamycin. Meanwhile, calpain-overexpressing capillary-like ECs were observed in the livers of high-fat diet–fed mice. Conditional KO of EC/hematopoietic Capns1, which encodes a calpain regulatory subunit, diminished levels of branched-chain amino acids in the hepatic microenvironment without altering plasma amino acid levels. Concomitantly, conditional KO of Capns1 mitigated hepatic steatosis without normalizing body weight and the plasma lipoprotein profile in an amino acid transporter–dependent manner. Mice with targeted Capns1 KO exhibited reduced phosphorylation of S6K and maturation of lipogenic factor sterol regulatory element–binding protein 1 in hepatocytes. Finally, we show that bone marrow transplantation negated the contribution of hematopoietic calpain systems. We conclude that overactivation of calpain systems may be responsible for the production of free amino acids in ECs, which may be sufficient to potentiate S6K/sterol regulatory element–binding protein 1–induced lipogenesis in surrounding hepatocytes.

Effect of wet-curing on physical properties and proteins of cured ham

AbstractConsuming enough protein is a very important part of a balanced and healthy diet. Complete proteins are those in which all essential amino acids are present. In meat products, like hams, many different reactions occur during ageing and storage. For example, the production of free amino acids or the production of biogenic amines is formed by decarboxylation. In this study, the presence of these amino acids and biogenic amines, as well as the quality properties of cured hams during curing and ageing, were investigated.The meat samples were immersed into 100 g L−1 NaCl brine. The curing took 20 days, followed by smoking and ageing for 35 days (12 °C, 75% RH). The wet-cured ham samples were analysed for changes in NaCl concentration (at 3 parts: surface, core, bottom layer). Moreover, color, water activity, denaturation temperature and enthalpy (Differential Scanning Calorimetry), free amino acids (FAAs) and biogenic amines (BAs) were also evaluated.