Pub Date : 2026-10-20DOI: 10.1021/acs.jafc.6c02612
Qi Zhang,Luyao Tang,Yanbo Zhao,Zhengxin Li,Yanpeng Ran,Zaigao Tan,Haifeng Chen,Yan Feng,Li Cui
The lipase enzyme shows great potential for catalyzing the synthesis of valuable food flavor compounds, such as γ- and δ-lactones, but natural lipases are limited by low conversion rates. In this study, we developed a computational-guided strategy named Synergistic Pairwise Residues Evolution for the efficient directed evolution of Candida antarctica lipase B (CalB). This approach identifies synergistic hotspot residues by combining substrate-contact information with evolutionarily coupled functional sectors. E188Q/I189F, the top-performing mutant with 2.75-fold improved activity, achieved over 99.5% conversion within 5 h at 37 °C, compared with only 49% for wild-type CalB in the synthesis of δ-dodecalactone. This study presents an efficient strategy for building functional enzyme toolkits and demonstrates their high catalytic performance in lactone synthesis.
{"title":"Synergistic Pairwise Residues Identification for Efficient Evolution of Candida antarctica Lipase B for Synthesis of γ- and δ-Lactones","authors":"Qi Zhang,Luyao Tang,Yanbo Zhao,Zhengxin Li,Yanpeng Ran,Zaigao Tan,Haifeng Chen,Yan Feng,Li Cui","doi":"10.1021/acs.jafc.6c02612","DOIUrl":"https://doi.org/10.1021/acs.jafc.6c02612","url":null,"abstract":"The lipase enzyme shows great potential for catalyzing the synthesis of valuable food flavor compounds, such as γ- and δ-lactones, but natural lipases are limited by low conversion rates. In this study, we developed a computational-guided strategy named Synergistic Pairwise Residues Evolution for the efficient directed evolution of Candida antarctica lipase B (CalB). This approach identifies synergistic hotspot residues by combining substrate-contact information with evolutionarily coupled functional sectors. E188Q/I189F, the top-performing mutant with 2.75-fold improved activity, achieved over 99.5% conversion within 5 h at 37 °C, compared with only 49% for wild-type CalB in the synthesis of δ-dodecalactone. This study presents an efficient strategy for building functional enzyme toolkits and demonstrates their high catalytic performance in lactone synthesis.","PeriodicalId":41,"journal":{"name":"Journal of Agricultural and Food Chemistry","volume":"88 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149218818","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-10-20DOI: 10.1021/acs.jafc.6c09395
Dong Xu,Zhimin Guo,Gen Xia,Shanshan Ling,Guohui Zhang,Xiaoli Tian,Kangsheng Ma
Spodoptera frugiperda, an economically important agricultural pest, has developed high resistance to chlorantraniliprole, yet mechanisms beyond metabolic detoxification and target-site mutations remain unclear. Chemosensory proteins (CSPs) have emerged as novel contributors to insecticide resistance via direct sequestration. Here, the transcript abundance of 22 CSPs was evaluated in susceptible (SS) and chlorantraniliprole-resistant (CAP-RR) strains of S. frugiperda using transcriptomics and RT-qPCR. Results showed that CSP10 was notably upregulated in the CAP-RR strain and inducible by the insecticide exposure. RNAi-mediated silencing of CSP10 significantly increased larval susceptibility to chlorantraniliprole. Fluorescence binding assays confirmed strong affinity between CSP10 and chlorantraniliprole. Molecular docking and dynamics further revealed the binding interaction primarily via van der Waals forces. Site-directed mutagenesis coupled with computational alanine scanning revealed Phe6, Ile17, Tyr21, Val25, and Phe38 as critical residues for the interaction. Collectively, our findings establish CSP10 as a key mediator of chlorantraniliprole resistance, offering a promising target for resistance management strategies.
{"title":"Chemosensory Protein 10 (CSP10) Is Involved in Resistance to Chlorantraniliprole in Spodoptera frugiperda","authors":"Dong Xu,Zhimin Guo,Gen Xia,Shanshan Ling,Guohui Zhang,Xiaoli Tian,Kangsheng Ma","doi":"10.1021/acs.jafc.6c09395","DOIUrl":"https://doi.org/10.1021/acs.jafc.6c09395","url":null,"abstract":"Spodoptera frugiperda, an economically important agricultural pest, has developed high resistance to chlorantraniliprole, yet mechanisms beyond metabolic detoxification and target-site mutations remain unclear. Chemosensory proteins (CSPs) have emerged as novel contributors to insecticide resistance via direct sequestration. Here, the transcript abundance of 22 CSPs was evaluated in susceptible (SS) and chlorantraniliprole-resistant (CAP-RR) strains of S. frugiperda using transcriptomics and RT-qPCR. Results showed that CSP10 was notably upregulated in the CAP-RR strain and inducible by the insecticide exposure. RNAi-mediated silencing of CSP10 significantly increased larval susceptibility to chlorantraniliprole. Fluorescence binding assays confirmed strong affinity between CSP10 and chlorantraniliprole. Molecular docking and dynamics further revealed the binding interaction primarily via van der Waals forces. Site-directed mutagenesis coupled with computational alanine scanning revealed Phe6, Ile17, Tyr21, Val25, and Phe38 as critical residues for the interaction. Collectively, our findings establish CSP10 as a key mediator of chlorantraniliprole resistance, offering a promising target for resistance management strategies.","PeriodicalId":41,"journal":{"name":"Journal of Agricultural and Food Chemistry","volume":"73 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149221649","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
To address the susceptibility of protein fibrils to pepsin degradation, this study proposed a fibril-to-microgel hierarchical assembly strategy, where flexible whey protein isolate fibrils (WPIF) and rigid flaxseed protein isolate fibrils (FPIF) were in situ self-cross-linked into microgels via a W/O emulsion template, followed by capsaicin (Cap) loading. Both microgels exhibited high gastric stability and intestinal response release properties. Compared with WPIF-based microgels (WPIFM), FPIF-based microgels (FPIFM) exhibited a more sustained intestinal digestion profile, leading to a higher Cap bioavailability. In high-fat-diet-induced obese mice, Cap-loaded microgels (WPIFM/Cap and FPIFM/Cap) effectively prevented metabolic disorders, as evidenced by reduced lipid accumulation, hepatic steatosis, and pro-inflammatory cytokines, with FPIFM/Cap demonstrating the highest protective efficacy. Both microgels modulated gut microbiota homeostasis, with FPIFM/Cap uniquely enriching Akkermansia, potentially due to its high arginine content and its stable fibrillar structure. This work provides a proof of concept for upcycling agricultural processing byproduct proteins into high-performance biomaterials for mitigating metabolic diseases.
{"title":"Oral Protein Fibril Microgels with Gastric Stability and Intestinal Targeting Enhance Capsaicin Bioavailability for Effective Obesity Prevention","authors":"Hui Zhang,Yaru Cui,Kaiwen Wu,Xuefeng Yin,Xiangyu Liu,Jun Liu,Yansong Xue,Ziqin Ye,Fen Li,Xing Li,Yuan Li","doi":"10.1021/acs.jafc.6c12370","DOIUrl":"https://doi.org/10.1021/acs.jafc.6c12370","url":null,"abstract":"To address the susceptibility of protein fibrils to pepsin degradation, this study proposed a fibril-to-microgel hierarchical assembly strategy, where flexible whey protein isolate fibrils (WPIF) and rigid flaxseed protein isolate fibrils (FPIF) were in situ self-cross-linked into microgels via a W/O emulsion template, followed by capsaicin (Cap) loading. Both microgels exhibited high gastric stability and intestinal response release properties. Compared with WPIF-based microgels (WPIFM), FPIF-based microgels (FPIFM) exhibited a more sustained intestinal digestion profile, leading to a higher Cap bioavailability. In high-fat-diet-induced obese mice, Cap-loaded microgels (WPIFM/Cap and FPIFM/Cap) effectively prevented metabolic disorders, as evidenced by reduced lipid accumulation, hepatic steatosis, and pro-inflammatory cytokines, with FPIFM/Cap demonstrating the highest protective efficacy. Both microgels modulated gut microbiota homeostasis, with FPIFM/Cap uniquely enriching Akkermansia, potentially due to its high arginine content and its stable fibrillar structure. This work provides a proof of concept for upcycling agricultural processing byproduct proteins into high-performance biomaterials for mitigating metabolic diseases.","PeriodicalId":41,"journal":{"name":"Journal of Agricultural and Food Chemistry","volume":"497 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149221653","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-10-20DOI: 10.1021/acs.jafc.6c08337
Anna Gajda,Ewelina Nowacka−Kozak,Piotr Cybulski
Antibiotic analysis is an important aspect of pig farming due to residues in tissues and the spread of antimicrobial resistance. In vivo control of antimicrobials is particularly significant, especially for the most critical and widely administered drugs. Thus, the aim of this study was to investigate the concentrations of tulathromycin and enrofloxacin in oral fluid after pig treatment. The analysis was performed by an ultrahigh-performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) method. Both drugs were detected for 31 days after the end of the treatment. On the last day of sampling, the concentration of tulathromycin was 19.7 μg/L and that of enrofloxacin was 26.4 μg/L. Tulathromycin reached its peak concentration (2180 μg/L) on day 2, while the highest concentration of enrofloxacin (1250 μg/L) was recorded on day 1 postadministration. The obtained results confirm the feasibility of detecting the two tested compounds in oral fluid, demonstrating a new approach for antemortem antibiotic control in pigs.
{"title":"Porcine Oral Fluid as a Diagnostic Matrix for Tulathromycin and Enrofloxacin Residue Surveillance","authors":"Anna Gajda,Ewelina Nowacka−Kozak,Piotr Cybulski","doi":"10.1021/acs.jafc.6c08337","DOIUrl":"https://doi.org/10.1021/acs.jafc.6c08337","url":null,"abstract":"Antibiotic analysis is an important aspect of pig farming due to residues in tissues and the spread of antimicrobial resistance. In vivo control of antimicrobials is particularly significant, especially for the most critical and widely administered drugs. Thus, the aim of this study was to investigate the concentrations of tulathromycin and enrofloxacin in oral fluid after pig treatment. The analysis was performed by an ultrahigh-performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) method. Both drugs were detected for 31 days after the end of the treatment. On the last day of sampling, the concentration of tulathromycin was 19.7 μg/L and that of enrofloxacin was 26.4 μg/L. Tulathromycin reached its peak concentration (2180 μg/L) on day 2, while the highest concentration of enrofloxacin (1250 μg/L) was recorded on day 1 postadministration. The obtained results confirm the feasibility of detecting the two tested compounds in oral fluid, demonstrating a new approach for antemortem antibiotic control in pigs.","PeriodicalId":41,"journal":{"name":"Journal of Agricultural and Food Chemistry","volume":"16 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149218821","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Liulin Baijiu (LLB), a representative Feng-flavor Baijiu, is highly regarded for its distinctive aroma and empty cup aroma; yet its chemical underpinnings remain unclear. This study employed an intergraded sensomics strategy encompassing sensory evaluation, GC-MS, GC-O-MS, AEDA, OAV, aroma recombination and omission tests. Repeated serial headspace solid-phase microextraction was applied to enhance the enrichment of trace low-volatility odorants remaining in the cup after ethanol volatilization. Nine key odorants were confirmed from 52 aroma-active compounds unveiled in LLB. Twenty-eight aroma-active compounds were identified in the 30-min empty cup sample, with six compounds achieving the highest flavor dilution (FD) factor of 729, including ethyl hexanoate, phenethyl acetate, hexanoic acid, ethyl laurate, phenol, and p-cresol, which contributed fruity, floral, fatty, and phenolic notes to the empty cup aroma. Collectively, these findings provide a molecular-scale perspective on its dynamic sensory evolution during cup-emptying process and may facilitate quality control and aroma-oriented product development.
{"title":"Sensomics-Guided Characterization of Liulin Baijiu and Its Empty Cup Volatilome: Uncovering the Key Odorants Behind Persistence","authors":"Huayang Wei,Jingyi Du,Boyu Li,Xinli Meng,Zhaofeng Zhang,Dongrui Zhao,Hehe Li,Bowen Wang,Mengyao Liu,Baoguo Sun,Jinyuan Sun","doi":"10.1021/acs.jafc.6c10770","DOIUrl":"https://doi.org/10.1021/acs.jafc.6c10770","url":null,"abstract":"Liulin Baijiu (LLB), a representative Feng-flavor Baijiu, is highly regarded for its distinctive aroma and empty cup aroma; yet its chemical underpinnings remain unclear. This study employed an intergraded sensomics strategy encompassing sensory evaluation, GC-MS, GC-O-MS, AEDA, OAV, aroma recombination and omission tests. Repeated serial headspace solid-phase microextraction was applied to enhance the enrichment of trace low-volatility odorants remaining in the cup after ethanol volatilization. Nine key odorants were confirmed from 52 aroma-active compounds unveiled in LLB. Twenty-eight aroma-active compounds were identified in the 30-min empty cup sample, with six compounds achieving the highest flavor dilution (FD) factor of 729, including ethyl hexanoate, phenethyl acetate, hexanoic acid, ethyl laurate, phenol, and p-cresol, which contributed fruity, floral, fatty, and phenolic notes to the empty cup aroma. Collectively, these findings provide a molecular-scale perspective on its dynamic sensory evolution during cup-emptying process and may facilitate quality control and aroma-oriented product development.","PeriodicalId":41,"journal":{"name":"Journal of Agricultural and Food Chemistry","volume":"236 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149221640","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sugar accumulation in fleshy fruits is largely dependent on membrane-localized sugar transporters, among which SWEET proteins play important roles. Although ABCB transporters, a subclass of the ATP-binding cassette (ABC) superfamily, mediate the transport of diverse substrates and participate in various physiological processes in plants, their direct involvement in fruit sugar accumulation remains unclear. In this study, SlABCB13, an ABCB transporter in tomato, was characterized. Multiple in vitro and in vivo assays demonstrated that SlABCB13 physically interacts with SlSWEET14. SlABCB13 was predominantly expressed in fruits and localized to the plasma membrane. Heterologous expression in yeast showed that SlABCB13 transported fructose and glucose, but not sucrose. Genetic analysis revealed that CRISPR/Cas9-mediated knockout of SlABCB13 significantly increased fruit hexose, sucrose, and total sugar contents while reducing fruit weight. SlABCB13-overexpressing lines also demonstrated reduced fruit weight, accompanied by stage-specific alterations in sugar levels. Transcriptome analysis revealed distinct transcriptional reprogramming in SlABCB13 knockout and overexpression lines, characterized by downregulation of sugar transporter genes in the overexpression lines, consistent with feedback inhibition, and upregulation in the knockout lines, indicative of compensatory activation. These results identify SlABCB13 as a plant ABCB transporter that mediates hexose transport in yeast and as an important regulator of sugar homeostasis in tomato fruits through coordinated feedback and compensatory mechanisms. This study broadens the functional spectrum of ABCB transporters and highlights SlABCB13 as a potential target for molecular breeding to improve fruit quality.
{"title":"SlABCB13 Regulates Hexose and Sucrose Homeostasis during Tomato Fruit Development","authors":"Xiao Xie,Na Xie,Tian Qi,Pengfei Liu,Qiuhui Li,Hongyu Qiao,Wei Wang,Xinsheng Zhang","doi":"10.1021/acs.jafc.6c08747","DOIUrl":"https://doi.org/10.1021/acs.jafc.6c08747","url":null,"abstract":"Sugar accumulation in fleshy fruits is largely dependent on membrane-localized sugar transporters, among which SWEET proteins play important roles. Although ABCB transporters, a subclass of the ATP-binding cassette (ABC) superfamily, mediate the transport of diverse substrates and participate in various physiological processes in plants, their direct involvement in fruit sugar accumulation remains unclear. In this study, SlABCB13, an ABCB transporter in tomato, was characterized. Multiple in vitro and in vivo assays demonstrated that SlABCB13 physically interacts with SlSWEET14. SlABCB13 was predominantly expressed in fruits and localized to the plasma membrane. Heterologous expression in yeast showed that SlABCB13 transported fructose and glucose, but not sucrose. Genetic analysis revealed that CRISPR/Cas9-mediated knockout of SlABCB13 significantly increased fruit hexose, sucrose, and total sugar contents while reducing fruit weight. SlABCB13-overexpressing lines also demonstrated reduced fruit weight, accompanied by stage-specific alterations in sugar levels. Transcriptome analysis revealed distinct transcriptional reprogramming in SlABCB13 knockout and overexpression lines, characterized by downregulation of sugar transporter genes in the overexpression lines, consistent with feedback inhibition, and upregulation in the knockout lines, indicative of compensatory activation. These results identify SlABCB13 as a plant ABCB transporter that mediates hexose transport in yeast and as an important regulator of sugar homeostasis in tomato fruits through coordinated feedback and compensatory mechanisms. This study broadens the functional spectrum of ABCB transporters and highlights SlABCB13 as a potential target for molecular breeding to improve fruit quality.","PeriodicalId":41,"journal":{"name":"Journal of Agricultural and Food Chemistry","volume":"56 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149221637","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-10-19DOI: 10.1021/acs.jafc.6c10478
Abdulqader Al-Adeeb,Sahibzada Muhammad Aqeel,Hend Sadeq Al-Jaberi,Qiuya Gu,Waleed Al-Ansi,Qais Ali Al-Maqtari,Wen-Long Liu,Xiaobin Yu
Cu/Zn superoxide dismutase (Cu/Zn-SOD) is a key antioxidant enzyme that catalyzes superoxide radical dismutation. Here, an integrated computational-assisted, structure-guided engineering strategy was applied to improve Cu/Zn-SOD catalytic performance. Gln22 and Glu134 were identified as promising engineering sites, and saturation mutagenesis yielded Q22I and E134 M as the most beneficial single substitutions, with specific activities 1.55- and 2.39-fold higher than wild type (WT), respectively. Combining both substitutions generated M2 (Q22I/E134M), which showed the highest specific activity of 7121.0 U mg–1, corresponding to a 3.43-fold increase over WT. The optimal temperature of Q22I and M2 increased from 40 to 50 °C, and M2 also exhibited enhanced thermostability. Kinetic analysis showed that M2 had a 41.18% lower Km and a 2.95-fold higher Vmax than WT. Molecular docking suggested complementary effects of Q22I and E134 M on structural stability and active-site optimization, supporting improved catalytic performance. These findings provide a basis for engineering efficient, thermostable SODs.
Cu/Zn超氧化物歧化酶（Cu/Zn- sod）是催化超氧化物自由基歧化酶的关键抗氧化酶。本文采用综合计算辅助、结构导向的工程策略来提高Cu/Zn-SOD的催化性能。Gln22和Glu134被确定为有前景的工程位点，饱和诱变得到Q22I和E134 M为最有利的单代替换，比活性分别比野生型（WT）高1.55倍和2.39倍。Q22I/E134M的比活性最高，为7121.0 U mg-1，比WT提高了3.43倍。Q22I和M2的最适温度从40℃提高到50℃，M2的热稳定性也得到了提高。动力学分析表明，M2的Km比WT低41.18%，Vmax比WT高2.95倍。分子对接表明，Q22I和E134 M在结构稳定性和活性位点优化方面具有互补作用，支持催化性能的提高。这些发现为工程上高效、耐热的sod提供了基础。
{"title":"Structure-Guided Engineering of Cu/Zn Superoxide Dismutase for Enhanced Catalytic Efficiency and Thermostability","authors":"Abdulqader Al-Adeeb,Sahibzada Muhammad Aqeel,Hend Sadeq Al-Jaberi,Qiuya Gu,Waleed Al-Ansi,Qais Ali Al-Maqtari,Wen-Long Liu,Xiaobin Yu","doi":"10.1021/acs.jafc.6c10478","DOIUrl":"https://doi.org/10.1021/acs.jafc.6c10478","url":null,"abstract":"Cu/Zn superoxide dismutase (Cu/Zn-SOD) is a key antioxidant enzyme that catalyzes superoxide radical dismutation. Here, an integrated computational-assisted, structure-guided engineering strategy was applied to improve Cu/Zn-SOD catalytic performance. Gln22 and Glu134 were identified as promising engineering sites, and saturation mutagenesis yielded Q22I and E134 M as the most beneficial single substitutions, with specific activities 1.55- and 2.39-fold higher than wild type (WT), respectively. Combining both substitutions generated M2 (Q22I/E134M), which showed the highest specific activity of 7121.0 U mg–1, corresponding to a 3.43-fold increase over WT. The optimal temperature of Q22I and M2 increased from 40 to 50 °C, and M2 also exhibited enhanced thermostability. Kinetic analysis showed that M2 had a 41.18% lower Km and a 2.95-fold higher Vmax than WT. Molecular docking suggested complementary effects of Q22I and E134 M on structural stability and active-site optimization, supporting improved catalytic performance. These findings provide a basis for engineering efficient, thermostable SODs.","PeriodicalId":41,"journal":{"name":"Journal of Agricultural and Food Chemistry","volume":"1 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149218868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ergosterol, a nutritionally relevant fungal sterol and vitamin D2 precursor, is valuable for food and feed applications, but its overproduction in Saccharomyces cerevisiae is constrained by stringent sterol homeostasis, limited organelle capacity, and the coordination of gene-dense pathways. A multilevel engineering framework was established by relieving inhibitory regulation, enhancing acetyl-CoA supply, strengthening rate-limiting steps, expanding endoplasmic reticulum capacity, and rewiring ethanol assimilation. A bidirectional-terminator-based integration toolkit enabled programmable chromosomal integration, modular pathway refactoring, and tunable expression partitioning. Stimulated Raman scattering (SRS) microscopy provided label-free visualization of intracellular ergosterol distribution and storage at single-cell resolution, supporting sterol homeostasis optimization. Robustness-oriented engineering of central carbon metabolism, mitochondrial function, and epigenetic regulation enhanced cellular fitness and fermentation performance. The resulting strain produced 0.983 g/L ergosterol in shake-flask cultures and 4.39 g/L in a 5-L bioreactor, corresponding to 51.7 mg/g dry cell weight (DCW), demonstrating the potential for scalable ergosterol biomanufacturing.
{"title":"Multilevel Engineering of Saccharomyces cerevisiae for Scalable Production of Ergosterol as a Vitamin D2 Precursor","authors":"Wenqian Wei,Qihang Chen,Chen Sang,Song Gao,Qian He,Sha Xu,Weizhu Zeng,Jingwen Zhou","doi":"10.1021/acs.jafc.6c05333","DOIUrl":"https://doi.org/10.1021/acs.jafc.6c05333","url":null,"abstract":"Ergosterol, a nutritionally relevant fungal sterol and vitamin D2 precursor, is valuable for food and feed applications, but its overproduction in Saccharomyces cerevisiae is constrained by stringent sterol homeostasis, limited organelle capacity, and the coordination of gene-dense pathways. A multilevel engineering framework was established by relieving inhibitory regulation, enhancing acetyl-CoA supply, strengthening rate-limiting steps, expanding endoplasmic reticulum capacity, and rewiring ethanol assimilation. A bidirectional-terminator-based integration toolkit enabled programmable chromosomal integration, modular pathway refactoring, and tunable expression partitioning. Stimulated Raman scattering (SRS) microscopy provided label-free visualization of intracellular ergosterol distribution and storage at single-cell resolution, supporting sterol homeostasis optimization. Robustness-oriented engineering of central carbon metabolism, mitochondrial function, and epigenetic regulation enhanced cellular fitness and fermentation performance. The resulting strain produced 0.983 g/L ergosterol in shake-flask cultures and 4.39 g/L in a 5-L bioreactor, corresponding to 51.7 mg/g dry cell weight (DCW), demonstrating the potential for scalable ergosterol biomanufacturing.","PeriodicalId":41,"journal":{"name":"Journal of Agricultural and Food Chemistry","volume":"11 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149218876","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alliin (S-allyl-l-cysteine sulfoxide) is a valuable bioactive sulfur compound and an allicin precursor. However, its biocatalytic synthesis is limited by poor stereocontrol of an Fe2+/α-ketoglutarate-dependent dioxygenase (l-isoleucine dioxygenase, IDO) that converts S-allyl-l-cysteine (SAC) into alliin. Herein, we analyzed the catalytic mechanism of IDO and proposed that the dihedral orientation of the SAC sulfur atom relative to the Fe(IV)═O intermediate likely governs sulfoxide chirality. Based on this mechanistic insight, we identified residue T233 and the flexible N77–F98 entrance loop as key determinants of substrate orientation during sulfoxidation. The engineered variant IDOM5 exhibited substantially enhanced diastereomeric excess (de, 93.4%) and turnover compared with the wild-type enzyme (de, 80.2%). Molecular dynamics simulations confirmed that the two geometric parameters encode stereochemical outcome and reaction rate, respectively, validating our initial hypothesis. Gram-scale biocatalytic synthesis of highly diastereomerically enriched (+)-alliin was achieved (2.70 g·L–1·h–1, conversion 93.70%). This work establishes a robust enzymatic platform for stereoselective alliin synthesis.
{"title":"Engineering of an Fe2+/α-Ketoglutarate-Dependent Dioxygenase Enables Stereoselective and Efficient Sulfoxide Synthesis","authors":"Zhaolin Song,Dequan Liu,Fangyue Bi,Yuying Gao,Zhenkun Shi,Jian Cheng,Wei Cao,Qinggang Li,Yu Li,Yihan Liu,Fuping Lu,Fenghua Wang","doi":"10.1021/acs.jafc.6c07879","DOIUrl":"https://doi.org/10.1021/acs.jafc.6c07879","url":null,"abstract":"Alliin (S-allyl-l-cysteine sulfoxide) is a valuable bioactive sulfur compound and an allicin precursor. However, its biocatalytic synthesis is limited by poor stereocontrol of an Fe2+/α-ketoglutarate-dependent dioxygenase (l-isoleucine dioxygenase, IDO) that converts S-allyl-l-cysteine (SAC) into alliin. Herein, we analyzed the catalytic mechanism of IDO and proposed that the dihedral orientation of the SAC sulfur atom relative to the Fe(IV)═O intermediate likely governs sulfoxide chirality. Based on this mechanistic insight, we identified residue T233 and the flexible N77–F98 entrance loop as key determinants of substrate orientation during sulfoxidation. The engineered variant IDOM5 exhibited substantially enhanced diastereomeric excess (de, 93.4%) and turnover compared with the wild-type enzyme (de, 80.2%). Molecular dynamics simulations confirmed that the two geometric parameters encode stereochemical outcome and reaction rate, respectively, validating our initial hypothesis. Gram-scale biocatalytic synthesis of highly diastereomerically enriched (+)-alliin was achieved (2.70 g·L–1·h–1, conversion 93.70%). This work establishes a robust enzymatic platform for stereoselective alliin synthesis.","PeriodicalId":41,"journal":{"name":"Journal of Agricultural and Food Chemistry","volume":"56 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149218881","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mycotoxins are common contaminants in agricultural products and feed, which pose a serious threat to public health. Zearalenone (ZEA) is a well-known environmental endocrine disruptor that causes reproductive disorders. Lycopene (LYC) is a possible therapeutic and preventive agent for male diseases. Here, we aim to explore the mechanism of ZEA-induced swine testicular (ST) cell injury and antagonism of lycopene. ST cells were treated with lycopene and/or ZEA for mechanistic validation. In this study, ZEA caused pronounced mitochondrial structural damage, whereas lycopene restored mitochondrial integrity. Lycopene also reduced ZEA-induced lipid peroxidation and ferrous ion levels, eventually inhibiting ferroptosis. Mechanistically, lycopene recovered PPARγ expression and regulated AMPK/mTOR signaling, thereby facilitating TFRC turnover. Of note, PPARγ knockdown weakened the effects of lycopene on ferroptosis, whereas TFRC overexpression diminished the protection afforded by autophagy activation. These findings indicate that lycopene mitigates ZEA-induced ferroptosis by regulating the PPARγ-AMPK-mTOR pathway and thus promoting autophagy-mediated TFRC degradation.
{"title":"PPARγ Is Critical for Lycopene Antagonizing Zearalenone-Induced Ferroptosis by Promoting Autophagy-Mediated TFRC Degradation in Swine Testicular Cells","authors":"Xin-Yue Mao,Zi-Yan Hu,Xin Yao,Ming Lou,Yue Cheng,Yi-Feng Huang,Hong-Li Si,Ming-Shan Chen,Jia-Xin Wang,Fu-Wei Jiang,Zhuo-Yu Liu,Xiao-Wei Li,Jin-Long Li,Yi Zhao","doi":"10.1021/acs.jafc.6c11442","DOIUrl":"https://doi.org/10.1021/acs.jafc.6c11442","url":null,"abstract":"Mycotoxins are common contaminants in agricultural products and feed, which pose a serious threat to public health. Zearalenone (ZEA) is a well-known environmental endocrine disruptor that causes reproductive disorders. Lycopene (LYC) is a possible therapeutic and preventive agent for male diseases. Here, we aim to explore the mechanism of ZEA-induced swine testicular (ST) cell injury and antagonism of lycopene. ST cells were treated with lycopene and/or ZEA for mechanistic validation. In this study, ZEA caused pronounced mitochondrial structural damage, whereas lycopene restored mitochondrial integrity. Lycopene also reduced ZEA-induced lipid peroxidation and ferrous ion levels, eventually inhibiting ferroptosis. Mechanistically, lycopene recovered PPARγ expression and regulated AMPK/mTOR signaling, thereby facilitating TFRC turnover. Of note, PPARγ knockdown weakened the effects of lycopene on ferroptosis, whereas TFRC overexpression diminished the protection afforded by autophagy activation. These findings indicate that lycopene mitigates ZEA-induced ferroptosis by regulating the PPARγ-AMPK-mTOR pathway and thus promoting autophagy-mediated TFRC degradation.","PeriodicalId":41,"journal":{"name":"Journal of Agricultural and Food Chemistry","volume":"234 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149218817","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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