Low-frequency alternating magnetic field and CaCl2 influence the physicochemical, conformational and gel characteristics of low-salt myofibrillar protein.

In this study, the improvement mechanism of low-frequency alternating magnetic field (LF-AMF, 5 mT, 3 h) combined with calcium chloride (CaCl2, 0-100 mM) on the gel characteristics of low-salt myofibrillar protein (MP) was investigated. LF-AMF combined with 80 mM CaCl2 treatment increased solubility (32.71%), surface hydrophobicity (40.86 µg), active sulfhydryl content (22.57%), water-holding capacity (7.15%). Besides, the combined treatment decreased turbidity, particle size and intrinsic fluorescence strength of MP. Fourier transform infrared spectroscopy (FT-IR) results indicated that the combined treatment altered the secondary structure of MP by increasing ß-sheet and ß-turn, and reducing α-helix and random coil. The combined treatment also induced a high G' value and shortened T2 relaxation time for forming a homogeneous and compact gel structure. These results revealed that LF-AMF combined CaCl2 treatment could as a potential approach for modifying the gel characteristics of low-salt MP.

Effect of low-frequency alternating magnetic fields on the physicochemical, conformational and rheological properties of myofibrillar protein after iterative freeze-thaw cycles.

In this work, the effects of low-frequency alternating magnetic fields (LF-AMF) on the physicochemical, conformational, and functional characteristics of myofibrillar protein (MP) after iterative freeze-thaw (FT) cycles were explored. With the increasing LF-AMF treatment time, the solubility, active sulfhydryl groups, surface hydrophobicity, emulsifiability, and emulsion stability of MP after five FT cycles evidently elevated and then declined, and the peak value was obtained at 3 h. Conversely, the moderate LF-AMF treatment time can significantly reduce the average particle size, carbonyl content, and endogenous fluorescence intensity of MP. The rheology results showed that various LF-AMF treatment times would elevate the G' value of MP after iterative FT cycles. The FTIR spectroscopy results suggested that LF-AMF influenced the secondary structure of MP after multiple FT cycles, resulting in a depression in α-helix content and an increment in ß-folding proportion. Moreover, LF-AMF treatment induced the gradually lighter and wider myosin heavy chain bands of MP, implying that LF-AMF accelerated the degradation of macromolecular aggregates. Therefore, the LF-AMF treatment efficaciously ameliorates the structural and functional deterioration of MP after iterative FT cycles and could be used as a potential quality-improving technology in the frozen meat industry.

Low-frequency alternating magnetic field thawing of frozen pork meat: Effects of intensity on quality properties and microstructure of meat and structure of myofibrillar proteins.

The purpose of the study was to evaluate the changes in quality properties and microstructure of pork meat as well as structural variation in myofibrillar proteins (MPs) after low-frequency alternating magnetic field thawing (LF-MFT) with different intensities (1-5 mT). LF-MFT at 3-5 mT shortened the thawing time. LF-MFT treatment significantly influenced the quality properties of meat and notably improved the structure of MPs (P < 0.05), compared to atmosphere thawing (AT). Especially, among the thawing treatments, LF-MFT-4 (LF-MFT at 4 mT) had the lowest values of thawing loss and drip loss, and the least changes in the color and myoglobin content. Regarding the results of rheological properties and micrographs, an optimal gel structure and a more compact muscle fiber arrangement formed during LF-MFT-4. Moreover, LF-MFT-4 was beneficial for improving the conformation of MPs. Therefore, LF-MFT-4 reduced the deterioration of porcine quality by protecting MPs structure, indicating a potential use in the meat thawing industry.

The effects of modified quinoa protein emulsion as fat substitutes in frankfurters.

In the current study, the effects of replacing different proportions of pork back fat (0, 25, 50, 75, and 100%) with high-pressure homogenization-modified quinoa protein emulsions (HMQE) on the physicochemical, water distribution, and rheological properties of low-fat frankfurters were investigated. HMQE incorporation significantly increased the moisture, ash, protein content, pH and L⁎ values and decreased the a⁎ and b⁎ values and T2 relaxation time of the low-fat frankfurters (P < 0.05). Of note, 50% fat substitution by HMQE yielded higher WHC, textural properties, gel strength, immobilized water percentage and G' value of the frankfurters than others. HMQE incorporation changed the protein secondary structure from α-helix to ß-sheet, forming a compact and uniform gel network structure with small cavities. Moreover, 50% fat substitution by HMQE did not affect sensory characteristics and improved the fat oxidative stability during storage. Therefore, the incorporation of HQME as a partial fat substitute resulted in nutritional benefits and quality enhancements, indicating that HQME could serve as a promising fat alternative in manufacturing low-fat frankfurters with desirable attributes.

Variation in glucosinolates and the formation of functional degradation products in two Brassica species during spontaneous fermentation.

Vegetables from the Brassica species are excellent sources of glucosinolates (GLSs), the precursors of health-promoting isothiocyanates (ITCs). Fermentation enhances the biotransformation of GLSs into potential bioactive ITCs. To explore the biotransformation of GLSs during Brassica fermentation, the changes in GLSs during the fermentation of two Brassica species (i.e., cauliflower and broccoli); the formation of corresponding breakdown products; and the shifts in physicochemical parameters, bacterial communities, and myrosinase activities involved in GLSs degradation were systematically investigated. Nine aliphatic, three indolic, and two benzenic GLSs were identified in fermented cauliflower (FC) and fermented broccoli (FB). Aliphatic glucoiberin and glucoraphanin were the major forms of GLS in FC and FB, respectively; indolic glucobrassicin was also abundant in both FC and FB. The total GLS content decreased by 85.29% and 65.48% after 3 d of fermentation in FC and FB, respectively. After 2 d of fermentation, a significant increase in bioactive GLS degradation products (P < 0.05), including sulforaphane (SFN), iberin (IBN), 3,3-diindolylmethane (DIM), and ascorbigen (ARG), was observed in FC and FB compared to in fresh cauliflower and broccoli. Moreover, variations in pH value and titratable acidity in FC and FB correlated with Brassica fermentation and were accomplished by lactic acid bacteria, including Weissella, Lactobacillus-related genera, Leuconostoc, Lactococcus, and Streptococcus. These changes may enhance the biotransformation of GSLs to ITCs. Overall, our results indicate fermentation leads to the degradation of GLSs and the accumulation of functional degradation products in FC and FB.

Effect of Artemisia sphaerocephala krasch gum on the functional properties of pork batters.

The influence of the incorporation of Artemisia sphaerocephala krasch gum (ASK gum; 0-0.18%) on the water holding capability (WHC), texture, color, rheological property, water distribution, protein conformation and microstructure of pork batters was investigated. The results showed that the cooking yield, WHC and L* value of pork batter gels significantly increased (p < .05) with the increasing incorporation of ASK gum, and the highest value were observed at 0.15%; the a* value decreased significantly (p < .05) and no significance was obtained in b* value (p > .05); the hardness, elasticity, cohesiveness and chewiness increased first and then decreased, and reached the highest value at 0.15%. The rheological results showed that the higher G' value was obtained in pork batters by the incorporation of ASK gum; the low field NMR analysis indicated that ASK gum significantly increased the proportion of P2b and P21 (p < .05) and decreased the proportion of P22 ; Fourier transform infrared spectroscopy (FTIR) indicated that the ASK gum significantly reduced the α-helix content and increased the ß-sheet content (p < .05). Scanning electron microscopy results suggested that the incorporation of ASK gum could promote the formation of a more homogeneous and stable microstructure of pork batter gels. Therefore, appropriate incorporation (0.15%) of ASK gum may improve the gel properties of pork batters, and while excessive incorporation (0.18%) could weaken the gel properties.

Understanding the influence of fluctuated low-temperature combined with high-humidity thawing on gelling properties of pork myofibrillar proteins.

The present study further investigated the effects of fluctuated low-temperature combined with high-humidity thawing (FLHT) on the gelling properties of pork myofibrillar proteins (MPs). Results showed that compared with refrigerator thawing (RT) and low-temperature combined with high-humidity thawing (LHT), FLHT effectively reduced the protein aggregation and maintained the relative stability by decreasing the variation in the turbidity and absolute ζ-potential value. The rheological results confirmed its improved elastic gel network. Meanwhile, FLHT samples exhibited markedly higher WHC with lower cooking loss (P < 0.05). The whiteness and strength of MPs gel were significantly higher in the FLHT group (P < 0.05). Moreover, there was no difference in textural properties between FLHT samples and fresh meat (FS) (P > 0.05), due to its homogeneous and compact microstructure. Therefore, FLHT plays an essential role in holding a superior gel quality and a compact structure, thereby evidencing its potential application in meat thawing.

High-pressure processing influences the conformation, water distribution, and gel properties of pork myofibrillar proteins containing Artemisia sphaerocephala Krasch gum.

The effect of high-pressure processing (100-4 00 MPa) on conformation, water distribution, and gel characteristics of reduced-sodium (0.3 M NaCl) myofibrillar protein containing 0.15% Artemisia sphaerocephala Krasch gum (AG) was investigated. The addition of AG resulted in the increase of WHC, proportion of immobilized water, and gel strength. Then, the WHC, proportion of immobilized water, and gel strength peaked after 200 MPa treatment, attributed to increased solubilization and zeta potential of MP, decreased particle size of MP, exposure of intrinsic tryptophan residues and the partial transformation of α-helix into ß-sheet in MP. Moreover, 300 and 400 MPa induced decreases in surface hydrophobicity, solubility and storage modulus, resulting in the formation of loose and disordered gel structures with attenuated WHC. These results suggest that application of moderate HPP (200 MPa) combined with AG could provide a novel approach to improve the WHC and gelation properties of reduced-sodium meat products.

Nonlinear Machine Learning in Warfarin Dose Prediction: Insights from Contemporary Modelling Studies.

Objective: This study aimed to systematically assess the characteristics and risk of bias of previous studies that have investigated nonlinear machine learning algorithms for warfarin dose prediction. Methods: We systematically searched PubMed, Embase, Cochrane Library, Chinese National Knowledge Infrastructure (CNKI), China Biology Medicine (CBM), China Science and Technology Journal Database (VIP), and Wanfang Database up to March 2022. We assessed the general characteristics of the included studies with respect to the participants, predictors, model development, and model evaluation. The methodological quality of the studies was determined, and the risk of bias was evaluated using the Prediction model Risk of Bias Assessment Tool (PROBAST). Results: From a total of 8996 studies, 23 were assessed in this study, of which 23 (100%) were retrospective, and 11 studies focused on the Asian population. The most common demographic and clinical predictors were age (21/23, 91%), weight (17/23, 74%), height (12/23, 52%), and amiodarone combination (11/23, 48%), while CYP2C9 (14/23, 61%), VKORC1 (14/23, 61%), and CYP4F2 (5/23, 22%) were the most common genetic predictors. Of the included studies, the MAE ranged from 1.47 to 10.86 mg/week in model development studies, from 2.42 to 5.18 mg/week in model development with external validation (same data) studies, from 12.07 to 17.59 mg/week in model development with external validation (another data) studies, and from 4.40 to 4.84 mg/week in model external validation studies. All studies were evaluated as having a high risk of bias. Factors contributing to the risk of bias include inappropriate exclusion of participants (10/23, 43%), small sample size (15/23, 65%), poor handling of missing data (20/23, 87%), and incorrect method of selecting predictors (8/23, 35%). Conclusions: Most studies on nonlinear-machine-learning-based warfarin prediction models show poor methodological quality and have a high risk of bias. The analysis domain is the major contributor to the overall high risk of bias. External validity and model reproducibility are lacking in most studies. Future studies should focus on external validity, diminish risk of bias, and enhance real-world clinical relevance.

Effects of high-pressure processing on the functional properties of pork batters containing Artemisia sphaerocephala krasch gum.

Here, the effect of high-pressure conditions (0.1-400 MPa) on the water-loss, texture, gel strength, color, dynamic rheological property, and water migration of pork batters containing 0.1% (W/W) Artemisia sphaerocephala krasch gum (PB-AG) is studied. Results indicated that the cooking yield, water-holding capacity, texture, gel strength, L* values, and G' values increased with the increase in pressure (0.1-300 MPa) (p < 0.05). Dynamic rheological results (G') revealed that the thermal gelling ability of the PB-AG gel gradually increased with pressure (0.1-300 MPa). The minimum of T22 content was observed and the proportion of immobilized water decreased at 300 MPa by low-filed nuclear magnetic resonance. However, excessive high-pressure processing treatments (400 MPa) resulted in lower gel strength, WHC, texture, and G'. The scanning electron microscopy results shown that a denser network structure with small cavities was observed at 300 MPa. Therefore, moderate pressure treatment (≤300 MPa) may improve gelation properties of PB-AG gel, while excessive pressure treatment (400 MPa) may weaken the gelation properties. PRACTICAL APPLICATION: High-pressure processing combining Artemisia sphaerocephala krasch gum could enhance the gelation properties of pork batters. To do so, establishing knowledge on gelation properties of pork batters with Artemisia sphaerocephala krasch gum at different pressure levels treatment would be of paramount importance, because this contributes furnishing engineering data pertinent to the technical progress for the processing of emulsion-type meat with high quality.

The effects of sodium chloride on proteins aggregation, conformation and gel properties of pork myofibrillar protein Running Head: Relationship aggregation, conformation and gel properties.

The objective of this study was to evaluate relationship with aggregation, secondary structures and gel properties of pork myofibrillar protein with different sodium chloride (1%, 2% and 3%). When the sodium chloride increased from 1 to 3%, the active sulfhydryl, surface hydrophobicity, hardness and cooking yield of myofibrillar protein were increased significantly (p < 0.05), the particle size, total sulfhydryl and Zeta potential were decreased significantly (p < 0.05), these meant the aggregations of pork myofibrillar protein were decreased. The changes of proteins aggregation induced the strongest intensity band of Amide I shifted up from 1660 cm-1 to 1661 cm-1, meanwhile, the ß-sheet structure content was increased significantly (p < 0.05) with the sodium chloride increased. From the above, the lower proteins aggregation and higher ß-sheet structure content could improve the water holding capacity and texture of pork myofibrillar protein gel.

Physicochemical and structural changes in myofibrillar proteins from porcine longissimus dorsi subjected to microwave combined with air convection thawing treatment.

The effect of microwave combined with air convection thawing (MAT) on the properties and tertiary structure of myofibrillar proteins (MPs) from porcine longissimus dorsi muscle was investigated and compared with single treatments (air thawing, microwave thawing) and fresh meat (FM). Among the thawing treatments, the carbonyl content, dityrosine content, and surface hydrophobicity of MPs in MAT were the lowest, whereas the total sulfhydryl content, water-holding capacity, and Ca2+-ATPase activity were the highest, suggesting that MAT retained MPs properties better. MAT possessed a more stable tertiary structure and exhibited slight changes in MPs aggregation and degradation. There was an insignificant difference (P > 0.05) in the immobilized water and free water between the MAT samples and FM, indicating a tighter interaction between water and muscle protein in MAT. Thus, MAT could retain the physicochemical and structural properties of MPs, which provided a combination of thawing treatments for application in meat industry.

Dynamics and dietary risk assessment of thiamethoxam in wheat, lettuce and tomato using field experiments and computational simulation.

Thiamethoxam is a widely used pesticide applied to different field crops. To inform risk assessment for this pesticide across relevant crops, we usually rely on field trials, which require time, costs and energy. For providing reliable data across crops and reduce experimental efforts, field trials should be complemented with dynamic modelling. In the present work, we hence focused on combining field trials with dynamic modelling to simulate mass evolutions of the pesticide-plant-system for thiamethoxam applied to wheat, lettuce and tomato as three major food crops. Field trials were conducted with QuEChERS (quick, easy, cheap, effective, rugged and safe) liquid chromatography-mass spectrometry, which gave consistent maximum residue concentrations for thiamethoxam in wheat, lettuce and tomato. We used these residues to evaluate the related dietary risk of humans consuming these food crops. Our results indicated that thiamethoxam did not provide any unacceptable dietary risk for humans across these three food crops, which is in line with findings from previous studies. Results for the studied crops could be extrapolated to other crops and with that, our study constitutes a cost- and time-efficient way of providing reliable input for risk assessment of pesticides across crops, which is relevant for both practitioners and regulators.

Physicochemical Properties and Protein Denaturation of Pork Longissimus Dorsi Muscle Subjected to Six Microwave-Based Thawing Methods.

Physicochemical changes and protein denaturation were evaluated for pork longissimus dorsi muscle subjected to different thawing methods. Fresh pork longissimus dorsi muscle served as a control. Microwave (MT), microwave combined with ultrasonic (MUT), microwave combined with 35 °C water immersion (MIT), microwave combined with 4 °C refrigeration (MRT), microwave combined with air convection (MAT), and microwave combined with running water (MWT) were applied. All microwave-based methods excepted for MT avoided localized overheating. The changes in the water holding capacity (WHC), color, TBARS, and protein solubility were lowest with MAT. Differential scanning calorimetry (DSC) and dynamic rheological property measurements indicated, that the MAT samples changed only slightly and presented with complete peaks and high G' values compared with the other treatments. Thus, MAT may reduce protein denaturation associated with meat thawing. The results of this study indicated that MAT effectively shortens thawing time, preserves meat quality and uniformity, and could benefit the meat industry and those who consume its products.

Residue behavior and dietary risk assessment of chlorothalonil and its metabolite SDS-3701 in water spinach to propose maximum residue limit (MRL).

Dietary risk assessment generally combines food consumption data and the concentration of pesticide by using the risk quotient (RQ) method. Chlorothalonil is the second popular fungicide in the world, and its residue and risk assessment in water spinach remain unknown. In this paper, the field trials of chlorothalonil in water spinach were operated under good agricultural practice (GAP) in China to human health protective. The dissipation experiments demonstrated that chlorothalonil was rapidly degraded in water spinach, with the half-lives of 1.8-3.2 days, and the amount of its metabolite SDS-3701 (4-hydroxy-2,5,6-trichloroisophthalonitrile) taken up through the water spinach roots from the soil was minor. The terminal experiments disclosed that the average residues of chlorothalonil and SDS-3701 in water spinach were below 6.59 mg/kg and 0.01 mg/kg, respectively. The results suggested the chronic dietary risk probability of chlorothalonil was 51.95-59.15% in terms of all registered crops, and the acute dietary risk probability of chlorothalonil was 12.30%-63.01% in water spinach, highlighting that the dietary risk of chlorothalonil in water spinach under GAP was acceptable. MRL of chlorothalonil was proposed as 7 mg/kg for water spinach and 5 days was recommended as a safe pre-harvest interval (PHI) for chlorothalonil application in water spinach field.

Dissipation behavior and dietary risk assessment of lambda-cyhalothrin, thiamethoxam and its metabolite clothianidin in apple after open field application.

The dissipation dynamics and residue amounts of lambda-cyhalothrin, thiamethoxam and clothianidin in apple were investigated by using rapid resolution liquid chromatography triple quadrupole mass spectrometer (RRLC-MS/MS) and gas chromatography mass spectrometry (GC-MS). The developed method performed satisfactory recoveries of 88%-105% and the limit of quantitation (LOQ) was 0.01 mg kg-1. The suspension concentrate (SC) formulation of lambda-cyhalothrin and thiamethoxam was applied on apple field in accordance with good agricultural practice (GAP). The half-lives of two pesticides ranged from 7.01 d to 17.3 d and the terminal residues were <0.01-0.21 mg kg-1. Based on the Chinese dietary pattern, the dietary risk of lambda-cyhalothrin and total thiamethoxam were predicted by comparing intake amounts with the toxicological data, namely acceptable daily intake (ADI) and acute reference dose (ARfD). The chronic and acute risk quotients were 0.1080-0.4463 and 0.0008-0.2005, respectively, which showed negligible risk for general consumers. The pre-harvest interval (PHI) of 21 d was suggested for the formulation in compliance with maximum residue limit (MRL) and dietary risk assessment, meanwhile, the MRL of 0.1 mg kg-1 was recommended for thiamethoxam in apple. These results were vital for guiding reasonable usage of two insecticides and for approval of formulation use.

A new regulator of cellulase and xylanase in the thermophilic fungus Myceliophthora thermophila strain ATCC 42464.

Myceliophthora thermophila (ATCC 42464) is a thermophilic fungus that produces cellulolytic enzymes with high thermal stability. Unlike its mesophile counterparts, study on gene expression regulation of cellulolytic enzymes in M. thermophila is inadequate. This work identified the function of MHR1, a putative transcription regulator of cellulolytic enzymes in M. thermophila that was found through RNA-Seq based gene expression profile analysis. RNA interference was used to study the role of MHR1. A recombinant plasmid, pUC19-Ppdc-mhr1-Tpdc, which contained the RNAi sequence for mhr1 was constructed and transformed into M. thermophila. One of the transformants, MtR5, in which the RNA interference efficiency was the highest, was used for the following studies. In the mhr1-silenced strain MtR5, the filter paper hydrolyzing activity was 1.33-fold; ß-1, 4-endoglucanase activity was 1.65-fold; and xylanase activity was 1.48-fold higher than those of the parental strain after induction, respectively, by wheat straw powder. qRT-PCR showed that gene expression of cbh1, cbh2, egl3 and xyr1 were 9.56-, 37.36-, 56.14- and 28.30-fold higher in MtR5 than in wild type, respectively. Our findings suggest that the transcription factor MHR1 of M. thermophila can repress cellulase and xylanase activities. Silenced mhr1 results in increased expression not only of the main cellulase genes, but also of the positive regulatory gene xyr1. This work is relevant to the development of M. thermophila as an industrial production host for cellulolytic and hemicellulolytic enzymes, which could be used to degrade a wide range of different biomass, converting lignocellulosic feedstock into sugar precursors for biofuels.

Mechanism of action of AMP-jsa9, a LI-F-type antimicrobial peptide produced by Paenibacillus polymyxa JSa-9, against Fusarium moniliforme.

LI-F type peptides (AMP-jsa9) are a group of cyclic lipodepsipeptides that exhibit broad antimicrobial spectrum against Gram-positive bacteria and filamentous fungi. We sought to assess the toxicity of AMP-jsa9 and the mechanism of AMP-jsa9 action against Fusarium moniliforme. AMP-jsa9 exhibited weak hemolytic activity and weak cytotoxicity at antimicrobial concentrations (32µg/ml). Confocal laser microscopy, SEM, and TEM indicated that AMP-jsa9 primarily targets the cell wall, plasma membrane, and cytoskeleton, increases membranepermeability, and enhances cytoplasm leakage (e.g., K+, protein). Quantitative proteomic analysis using isobaric tags for relative and absolute quantitation (iTRAQ) detected a total of 162 differentially expressed proteins (59 up-regulated and 103 down-regulated) following treatment of F. moniliforme with AMP-jsa9. AMP-jsa9 treatment also led to reductions in chitin, ergosterol, NADH, NADPH, and ATP levels. Moreover, fumonisin B1 expression and biosynthesis was suppressed in AMP-jsa9-treated F. moniliforme. Our results provide a theoretical basis for the application of AMP-jsa9 as a natural and effective antifungal agent in the agricultural, food, and animal feed industries.

iTRAQ-based proteomic analysis of LI-F type peptides produced by Paenibacillus polymyxa JSa-9 mode of action against Bacillus cereus.

LI-F type peptides (AMP-jsa9) produced by Paenibacillus polymyxa JSa-9 are a group of cyclic lipodepsipeptide antibiotics that exhibit a broad antimicrobial spectrum against Gram-positive bacteria and filamentous fungi, especially Bacillus cereus and Fusarium moniliforme. In this study, to better understand the antibacterial mechanism of AMP-jsa9 against B. cereus, the ultrastructure of AMP-jsa9-treated B. cereus cells was observed by both atomic force microscopy and transmission electron microscopy, and quantitative proteomic analysis was performed on proteins extracted from treated and untreated bacterial cells by using isobaric tag for relative and absolute quantitation (iTRAQ) labeling and LC-MS/MS analysis to access differentially expressed proteins. Furthermore, multiple experiments were conducted to validate the results of the proteomic analysis, including determinations of ATP, NAD(+)H, NADP(+)H, reactive oxygen species (ROS), the activities of catalase (CAT) and superoxide dismutase (SOD), and the relative expression of target genes by quantitative real-time PCR. Bacterial cells exposed to AMP-jsa9 showed irregular surfaces with bleb projections and concaves; we hypothesize that AMP-jsa9 penetrated the cell wall and was anchored on the cytoplasmic membrane and that ROS accumulated in the cell membrane after treatment with AMP-jsa9, modulating the bacterial membrane properties and increasing membrane permeability. Consequently, the blebs were formed on the cell wall by the impulsive force of the leakage of intercellular contents. iTRAQ-based proteomic analysis detected a total of 1317 proteins, including 176 differentially expressed proteins (75 upregulated (fold >2) and 101 downregulated (fold <0.5)). Based on proteome analysis, the putative pathways of AMP-jsa9 action against B. cereus can be summarized as: (i) inhibition of bacterial sporulation, thiamine biosynthesis, energy metabolism, DNA transcription and translation, and cell wall biosynthesis, through direct regulation of protein levels; and (ii) indirect effects on the same pathways through the accumulation of ROS and the consequent impairment of cellular functions, resulting from downregulation of antioxidant proteins, especially CAT and SOD. BIOLOGICAL SIGNIFICANCE: The mode of action of LI-F type antimicrobial peptides (AMP-jsa9) against B. cereus was elucidated at the proteomic level. Two pathways of AMP-jsa9 action upon B. cereus cells were identified and the mechanism of bleb formation on the surfaces of bacterial cells was predicted based on the results of ultrastructural observation and proteomic analysis. These results are helpful in understanding the mechanism of LI-F type peptides and in providing the theoretical base for applying AMP-jsa9 or its analogs to combat Gram-positive pathogenic bacteria in the food and feed industries.

Characterization of a Bacillus subtilis surfactin synthetase knockout and antimicrobial activity analysis.

Gene knockout is an important approach to improve the production of antimicrobial compounds. B. subtilis PB2-LS10, derived from B. subtilis PB2-L by a surfactin synthetase (srf) genes knockout, exhibits stronger inhibitory action than its parental strain against all tested pathogenic bacteria and fungi. The antimicrobial extracts produced by B. subtilis PB2-L and B. subtilis PB2-LS10 respectively were characterized by the high-resolution LC-ESI-MS. To provide further insight into the distinct antimicrobial activities, we investigated the impact of the srf genes deletion on the growth and gene transcriptional profile of the strains. The mutant strain grew quickly and reached stationary phase 2h earlier than the wild-type. Prominent expression changes in the modified strain involved genes that were essential to metabolic pathways and processes. Genes related to amino acid transport, ATP-binding cassette (ABC) transporters and protein export were up-regulated in strain PB2-LS10. However, amino acid metabolism, carbohydrate metabolism and fatty acid metabolism were repressed. Because of its excellent antimicrobial activity, strain PB2-LS10 has potential for use in food preservation.