Pub Date : 2025-02-01DOI: 10.1016/j.procbio.2024.12.013
Munirah Munirah , Dudi Hardianto , Efrida Martius , Uli Julia Nasution , Anna Safarrida
The synthesis of insulin using recombinant Pichia pastoris represents a significant advancement in biotechnology with profound implications for healthcare. This review explores the process of insulin production in P. pastoris. The selection of P. pastoris as a host organism for insulin production is discussed. The stages of insulin production, including gene cloning, vector construction, transformation, and expression, are outlined, emphasizing the importance of strain selection and optimization of fermentation conditions. The review also examines factors influencing insulin production, such as strain phenotype, fermentation media composition, and induction strategies. Additionally, the role of glycerol and methanol in fermentation for insulin production and the impact of temperature and pH on recombinant protein expression are elucidated. Overall, this review provides insights into the comprehensive approach required for achieving high-level insulin precursor production in P. pastoris, offering valuable guidance for further advancements in recombinant protein expression technology and healthcare biotechnology.
{"title":"Insulin production in Pichia pastoris: Mini-review of biotechnological advancements and process optimization","authors":"Munirah Munirah , Dudi Hardianto , Efrida Martius , Uli Julia Nasution , Anna Safarrida","doi":"10.1016/j.procbio.2024.12.013","DOIUrl":"10.1016/j.procbio.2024.12.013","url":null,"abstract":"<div><div>The synthesis of insulin using recombinant <em>Pichia pastoris</em> represents a significant advancement in biotechnology with profound implications for healthcare. This review explores the process of insulin production in <em>P. pastoris</em>. The selection of <em>P. pastoris</em> as a host organism for insulin production is discussed. The stages of insulin production, including gene cloning, vector construction, transformation, and expression, are outlined, emphasizing the importance of strain selection and optimization of fermentation conditions. The review also examines factors influencing insulin production, such as strain phenotype, fermentation media composition, and induction strategies. Additionally, the role of glycerol and methanol in fermentation for insulin production and the impact of temperature and pH on recombinant protein expression are elucidated. Overall, this review provides insights into the comprehensive approach required for achieving high-level insulin precursor production in <em>P. pastoris</em>, offering valuable guidance for further advancements in recombinant protein expression technology and healthcare biotechnology.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"149 ","pages":"Pages 277-287"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143139253","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.procbio.2024.11.033
Jun Young Choi , Jaepyeong Jang , Young-Chul Park , Pyung Cheon Lee
In this study, we engineered a synthetic promoter library for Lactobacillus casei BL23 by randomizing the −35, −10 regions, and spacer sequences of the aldolase promoter. The goal was to generate promoters with varying strengths for use in metabolic engineering applications. Using computational tools (SAPPHIRE and BacPP), we identified core promoter elements and systematically randomized these regions to create 31 variants. Fluorescence-activated cell sorting (FACS) allowed for the isolation of weak, moderate, and strong promoters based on superfolder GFP (sfGFP) expression levels. The strongest promoter exhibited an 8.71-fold increase in sfGFP expression compared to the native aldolase promoter. Sequence analysis revealed specific nucleotide preferences in the core elements, which influenced promoter strength. This study offers a valuable platform for fine-tuning gene expression in L. casei, providing insights for future metabolic engineering applications.
{"title":"Systematic promoter engineering in Lactobacillus casei: Construction and screening of a synthetic aldolase promoter library for enhanced gene expression","authors":"Jun Young Choi , Jaepyeong Jang , Young-Chul Park , Pyung Cheon Lee","doi":"10.1016/j.procbio.2024.11.033","DOIUrl":"10.1016/j.procbio.2024.11.033","url":null,"abstract":"<div><div>In this study, we engineered a synthetic promoter library for <em>Lactobacillus casei</em> BL23 by randomizing the −35, −10 regions, and spacer sequences of the aldolase promoter. The goal was to generate promoters with varying strengths for use in metabolic engineering applications. Using computational tools (SAPPHIRE and BacPP), we identified core promoter elements and systematically randomized these regions to create 31 variants. Fluorescence-activated cell sorting (FACS) allowed for the isolation of weak, moderate, and strong promoters based on superfolder GFP (sfGFP) expression levels. The strongest promoter exhibited an 8.71-fold increase in sfGFP expression compared to the native aldolase promoter. Sequence analysis revealed specific nucleotide preferences in the core elements, which influenced promoter strength. This study offers a valuable platform for fine-tuning gene expression in <em>L. casei</em>, providing insights for future metabolic engineering applications.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"149 ","pages":"Pages 45-53"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143139879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.procbio.2024.11.030
Min Qu, Jingbo Kang, Xiuqing Zhu, Xin Zhang, Linlin Liu, Ying Zhu, Yuyang Huang, Bingyu Sun, Mingshou Lu
Quorum sensing (QS) plays a key role in the spoilage of food spoilage bacteria, and inhibiting the spoilage phenotype by interfering with QS has become a hot research topic in recent years. Previous studies showed that Nisin had a wide range of targets and its bacteriostatic activity focused on inhibiting Gram-positive bacteria. In this paper, we investigated the effect of Nisin at sub-MIC on virulence factors and biofilm formation of the common food spoilage bacteria Bacillus licheniformis DSM13 and Bacillus amyloliquefaciens CC178. The results showed that the amount of biofilm formation, EPS production, protease activity and amylase activity, promoter expression, and motility of the two bacteria showed different degrees of reduction, but hardly affected the normal proliferation of the bacteria; The impaired biofilm integrity and a large amount of matrix reduction were observed by SEM; The molecular docking demonstrated that Nisin non-competitively bind to the com P protein-specific of the two bacteria through hydrogen bonding sites, affecting the binding of com X to com P. These results suggested that low concentrations of Nisin effectively reduced the risk and safety hazards caused by QS of G+ and is a promising QS inhibitor.
{"title":"Inhibition of virulence factors and biofilm formation of Bacillus licheniformis and Bacillus amyloliquefaciens by Nisin","authors":"Min Qu, Jingbo Kang, Xiuqing Zhu, Xin Zhang, Linlin Liu, Ying Zhu, Yuyang Huang, Bingyu Sun, Mingshou Lu","doi":"10.1016/j.procbio.2024.11.030","DOIUrl":"10.1016/j.procbio.2024.11.030","url":null,"abstract":"<div><div>Quorum sensing (QS) plays a key role in the spoilage of food spoilage bacteria, and inhibiting the spoilage phenotype by interfering with QS has become a hot research topic in recent years. Previous studies showed that Nisin had a wide range of targets and its bacteriostatic activity focused on inhibiting Gram-positive bacteria. In this paper, we investigated the effect of Nisin at sub-MIC on virulence factors and biofilm formation of the common food spoilage bacteria <em>Bacillus licheniformis</em> DSM13 and <em>Bacillus amyloliquefaciens</em> CC178. The results showed that the amount of biofilm formation, EPS production, protease activity and amylase activity, promoter expression, and motility of the two bacteria showed different degrees of reduction, but hardly affected the normal proliferation of the bacteria; The impaired biofilm integrity and a large amount of matrix reduction were observed by SEM; The molecular docking demonstrated that Nisin non-competitively bind to the com P protein-specific of the two bacteria through hydrogen bonding sites, affecting the binding of com X to com P. These results suggested that low concentrations of Nisin effectively reduced the risk and safety hazards caused by QS of G+ and is a promising QS inhibitor.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"149 ","pages":"Pages 54-64"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143139880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.procbio.2024.12.021
Puhong Yi , Yue Xu , Hanlin Liu , Yuhua Hao , Mengdan Liu , Zhiqiang Liu , Yaping Xue , Liqun Jin , Yuguo Zheng
2-oxo-4-[(hydroxy)(methyl)phosphinoyl]butyric acid (PPO) is a valuable prochiral keto acid for the asymmetric synthesis of the important herbicide L-phosphinothricin (L-PPT). Developing an efficient biocatalytic pathway to meet the increasing demand for PPO is highly attractive. In this study, a robust D-amino acid aminotransferase from Cytobacillus firmus (CfDAAT) was screened, which exhibited excellent tolerability to D,L-PPT and showed remarkable specific activity and affinity toward pyruvate (61.9 U/mg, Km = 7.4 mM). A designed co-substrate regeneration system was then introduced to shift the reaction equilibrium forward by coupling with D-amino acid oxidase (DAAO) and catalase (CAT). An efficient whole-cell biocatalyst was obtained by fine-tuning RBS intensity to coordinate the co-expression of the three enzymes. Moreover, the optimum conditions to promote overall conversion efficiency were identified by response surface methodology. Consequently, with only a catalytic amount of pyruvate (3.33 mol%) added in a 1 L reaction system, 200 mM D-PPT (400 mM D,L-PPT) was completely converted in 10 h, resulting in a PPO productivity of 3.33 g⋅L−1⋅h−1, and a notable conversion rate of > 99 % at 300 mM D-PPT in 16 h. It demonstrated a considerable catalytic level and provided a competitive route for industrial PPO production from D,L-PPT.
{"title":"Efficient kinetic resolution of D,L-phosphinothricin using an aminotransferase-mediated cascade","authors":"Puhong Yi , Yue Xu , Hanlin Liu , Yuhua Hao , Mengdan Liu , Zhiqiang Liu , Yaping Xue , Liqun Jin , Yuguo Zheng","doi":"10.1016/j.procbio.2024.12.021","DOIUrl":"10.1016/j.procbio.2024.12.021","url":null,"abstract":"<div><div>2-oxo-4-[(hydroxy)(methyl)phosphinoyl]butyric acid (PPO) is a valuable prochiral keto acid for the asymmetric synthesis of the important herbicide L-phosphinothricin (L-PPT). Developing an efficient biocatalytic pathway to meet the increasing demand for PPO is highly attractive. In this study, a robust D-amino acid aminotransferase from <em>Cytobacillus firmus</em> (<em>Cf</em>DAAT) was screened, which exhibited excellent tolerability to D,L-PPT and showed remarkable specific activity and affinity toward pyruvate (61.9 U/mg, Km = 7.4 mM). A designed co-substrate regeneration system was then introduced to shift the reaction equilibrium forward by coupling with D-amino acid oxidase (DAAO) and catalase (CAT). An efficient whole-cell biocatalyst was obtained by fine-tuning RBS intensity to coordinate the co-expression of the three enzymes. Moreover, the optimum conditions to promote overall conversion efficiency were identified by response surface methodology. Consequently, with only a catalytic amount of pyruvate (3.33 mol%) added in a 1 L reaction system, 200 mM D-PPT (400 mM D,L-PPT) was completely converted in 10 h, resulting in a PPO productivity of 3.33 g⋅L<sup>−1</sup>⋅h<sup>−1</sup>, and a notable conversion rate of > 99 % at 300 mM D-PPT in 16 h. It demonstrated a considerable catalytic level and provided a competitive route for industrial PPO production from D,L-PPT.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"149 ","pages":"Pages 295-305"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143139497","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
C-Phycocyanin (CPC) is a pigment protein produced by cyanobacteria and is commonly used as a food coloring and in cosmetics. For the separation and concentration of CPC, a novel combined process of chromatographic column and ultrafiltration module is proposed. The unit of the column and ultrafiltration module are described mathematically to calculate the combined units as a process. The ion-exchange resin-packed column separated CPC with a purity of 2.0 or higher at a rate of 15 mL per cycle. Ultrafiltration module (MWCO 30 kDa) was operated in desalting mode (diafiltration: DF) and concentration mode (ultrafiltration: UF). DF reduced the salt concentration from an initial concentration of 260 mM to 40 mM, with no proteins leaking from the module. UF increased CPC concentration from 0.82 g/L to 1.48 g/L. Parameters were determined using a mathematical model to simulate the combined process of ion-exchange resin-packed column and ultrafiltration module, and conditions (permeation velocity and time of ultrafiltration) were determined to achieve the desired product characteristics. Different from separation unit operation, combined process of a resin-packed column and ultrafiltration module was capable of separating CPC with approximately twice the productivity of other methods.
{"title":"Separation of C-Phycocyanin from Spirulina sp. using the coupled processes of an ion-exchange resin-packed column and ultrafiltration module","authors":"Takanori Hidane , Haruka Miyoshi , Mikihide Demura , Shintaro Morisada , Keisuke Ohto , Hidetaka Kawakita","doi":"10.1016/j.procbio.2024.12.020","DOIUrl":"10.1016/j.procbio.2024.12.020","url":null,"abstract":"<div><div>C-Phycocyanin (CPC) is a pigment protein produced by cyanobacteria and is commonly used as a food coloring and in cosmetics. For the separation and concentration of CPC, a novel combined process of chromatographic column and ultrafiltration module is proposed. The unit of the column and ultrafiltration module are described mathematically to calculate the combined units as a process. The ion-exchange resin-packed column separated CPC with a purity of 2.0 or higher at a rate of 15 mL per cycle. Ultrafiltration module (MWCO 30 kDa) was operated in desalting mode (diafiltration: DF) and concentration mode (ultrafiltration: UF). DF reduced the salt concentration from an initial concentration of 260 mM to 40 mM, with no proteins leaking from the module. UF increased CPC concentration from 0.82 g/L to 1.48 g/L. Parameters were determined using a mathematical model to simulate the combined process of ion-exchange resin-packed column and ultrafiltration module, and conditions (permeation velocity and time of ultrafiltration) were determined to achieve the desired product characteristics. Different from separation unit operation, combined process of a resin-packed column and ultrafiltration module was capable of separating CPC with approximately twice the productivity of other methods.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"149 ","pages":"Pages 260-269"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143139501","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.procbio.2024.11.036
Júlia Firme Freitas, Douglas Felipe de Lima Silva, Jenielly Noronha Ferreira Castro, Lucymara Fassarella Agnez-Lima
A recurring theme is the quest for microorganisms with a high capability to degrade toxic hydrocarbons or produce biosurfactants to aid in bioremediation. This study identified two new bacterial species isolated from samples collected in Brazilian oil reservoirs. The phylogenetic analysis revealed two distinct clusters. AP1BH01–1, BD18, BD22, BD23, and BD61 formed one cluster that resembles Ochrobactrum intermedium. In contrast, BD67 clustered more closely with Ochrobactrum pseudointermedium. The whole genome analysis showed significant genetic differences from known species within the Ochrobactrum genus. OrthoANI (< 91.14 %) and dDDH (< 42.80 %) values for our isolates were notably lower than expected for species classification and supported by genomic correlation matrix cluster, synteny, pan-genome, and phylogenetical analysis. Our isolates can utilize petroleum as a carbon source, showing unique genes dedicated to the degradation of aromatic hydrocarbons and metal detoxification. Notably, the isolates can degrade petroleum, pyrene, and hexacosene. Moreover, despite the absence of a cluster dedicated to biosurfactant production, all isolates demonstrated their proficiency in emulsifier production. The isolates exhibit distinctive genotypic and phenotypic characteristics compared to the known Ochrobactrum species. Considering these findings, we propose naming AP1BH01–1, BD18, BD22, BD23, and BD61 as Ochrobactrum oleinvorans and BD67 as Ochrobactrum oleophilic.
{"title":"Genomic and phenotypic characterization of novel Ochrobactrum species isolated from Brazilian oil reservoirs: Genomic diversity and bioremediation potential","authors":"Júlia Firme Freitas, Douglas Felipe de Lima Silva, Jenielly Noronha Ferreira Castro, Lucymara Fassarella Agnez-Lima","doi":"10.1016/j.procbio.2024.11.036","DOIUrl":"10.1016/j.procbio.2024.11.036","url":null,"abstract":"<div><div>A recurring theme is the quest for microorganisms with a high capability to degrade toxic hydrocarbons or produce biosurfactants to aid in bioremediation. This study identified two new bacterial species isolated from samples collected in Brazilian oil reservoirs. The phylogenetic analysis revealed two distinct clusters. AP1BH01–1, BD18, BD22, BD23, and BD61 formed one cluster that resembles <em>Ochrobactrum intermedium</em>. In contrast, BD67 clustered more closely with <em>Ochrobactrum pseudointermedium</em>. The whole genome analysis showed significant genetic differences from known species within the <em>Ochrobactrum</em> genus. OrthoANI (< 91.14 %) and dDDH (< 42.80 %) values for our isolates were notably lower than expected for species classification and supported by genomic correlation matrix cluster, synteny, pan-genome, and phylogenetical analysis. Our isolates can utilize petroleum as a carbon source, showing unique genes dedicated to the degradation of aromatic hydrocarbons and metal detoxification. Notably, the isolates can degrade petroleum, pyrene, and hexacosene. Moreover, despite the absence of a cluster dedicated to biosurfactant production, all isolates demonstrated their proficiency in emulsifier production. The isolates exhibit distinctive genotypic and phenotypic characteristics compared to the known <em>Ochrobactrum</em> species. Considering these findings, we propose naming AP1BH01–1, BD18, BD22, BD23, and BD61 as <em>Ochrobactrum oleinvorans</em> and BD67 as <em>Ochrobactrum oleophilic</em>.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"149 ","pages":"Pages 74-84"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143139504","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.procbio.2024.12.016
Zhuhui Yang , Yumei Han , Runping Yang , Ning Guo , Huizhen Duan , Guiqin Zhang
Two bio-trickling filters (BTFs) were set up for ethyl acetate removal: one of the filters contained a built-in proton exchange membrane (M-BTF), and the other contained the conventional structure (O-BTF). The comparative results showed that M-BTF had superior biodegradation capacity. Its start-up was relatively faster, and it produced more biomass and exhibited a higher removal efficiency, achieving a maximum removal efficiency difference of 9.9 % than O-BTF. Meanwhile, the maximum output voltage of 536 mV was obtained with the corresponding removal loading of 249.3 g•m−3•h−1. The intermediate product was detected to explore the possible degradation pathway of ethyl acetate, and results showed that the degradation of ethyl acetate was accompanied by the synthesis of long-chain hydrocarbons. The microbial analysis revealed that the anode electrode was highly selective to the bacterial strains, resulting in more electroactive bacteria containing Pseudomonas, Flavobacterium, Pseudoxanthomonas, and Geobacter constructing the microbial community in M-BTF.
{"title":"Enhancement of gaseous ethyl acetate removal in a new bio-trickling filter by integrating with microbial fuel cell","authors":"Zhuhui Yang , Yumei Han , Runping Yang , Ning Guo , Huizhen Duan , Guiqin Zhang","doi":"10.1016/j.procbio.2024.12.016","DOIUrl":"10.1016/j.procbio.2024.12.016","url":null,"abstract":"<div><div>Two bio-trickling filters (BTFs) were set up for ethyl acetate removal: one of the filters contained a built-in proton exchange membrane (M-BTF), and the other contained the conventional structure (O-BTF). The comparative results showed that M-BTF had superior biodegradation capacity. Its start-up was relatively faster, and it produced more biomass and exhibited a higher removal efficiency, achieving a maximum removal efficiency difference of 9.9 % than O-BTF. Meanwhile, the maximum output voltage of 536 mV was obtained with the corresponding removal loading of 249.3 g•m<sup>−3</sup>•h<sup>−1</sup>. The intermediate product was detected to explore the possible degradation pathway of ethyl acetate, and results showed that the degradation of ethyl acetate was accompanied by the synthesis of long-chain hydrocarbons. The microbial analysis revealed that the anode electrode was highly selective to the bacterial strains, resulting in more electroactive bacteria containing <em>Pseudomonas, Flavobacterium, Pseudoxanthomonas,</em> and <em>Geobacter</em> constructing the microbial community in M-BTF.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"149 ","pages":"Pages 270-276"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143139878","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.procbio.2024.12.007
Miroslava Kačániová , Mária Škultétyová , Eva Tvrdá , Filip Benko , Michal Ďuračka , Natália Čmiková , Eva Ivanišová , Jaroslav Havlík , Grzegorz Zaguła , Angel Antonio Carbonell Barrachina , Stefania Garzoli
The objective of this study was to compare the probiotic bacteria present in drone milk (DM) and drone larvae (DL). Furthermore, the study aimed to investigate the antibacterial capacity against Staphylococcus and Streptococcus species isolated from animal semen. The in vitro effects of DM and DL on male reproductive cells and rabbit tissues were examined. In addition, the nutritional and chemical composition of both bee products was investigated. Fresh and freeze-dried samples were used to isolate probiotic bacteria identified by MALDI-TOF MS Biotyper. Lactobacilli, especially Apilactobacillus kunkeii, Lactiplantibacillus pentosus, Lactiplantibacillus plantarum, and Secundilactobacillus malefermentas, received the highest identification scores. The best results were observed when targeting S. aureus, S. epidermidis, and S. vitulinus. The freeze-dried materials were tested on rabbit spermatozoa and testicular fragments to ascertain the effects of DM and DL. The results showed that the samples significantly reduced the amount of reactive oxygen species in the reproductive structures which resulted in a decrease in lipid peroxidation. Furthermore, testicular fragments treated in vitro with DM and DL produced considerably more testosterone and cholesterol; crude protein, nitrogenous compounds and fat were all present in higher amounts in DL while the content of reducing sugars was higher in DM.
{"title":"Nutritional value and chemical properties of drone milk as a source of probiotics and evaluation of antioxidant effects on reproductive structures by in vitro test","authors":"Miroslava Kačániová , Mária Škultétyová , Eva Tvrdá , Filip Benko , Michal Ďuračka , Natália Čmiková , Eva Ivanišová , Jaroslav Havlík , Grzegorz Zaguła , Angel Antonio Carbonell Barrachina , Stefania Garzoli","doi":"10.1016/j.procbio.2024.12.007","DOIUrl":"10.1016/j.procbio.2024.12.007","url":null,"abstract":"<div><div>The objective of this study was to compare the probiotic bacteria present in drone milk (DM) and drone larvae (DL). Furthermore, the study aimed to investigate the antibacterial capacity against <em>Staphylococcus</em> and <em>Streptococcus</em> species isolated from animal semen. The <em>in vitro</em> effects of DM and DL on male reproductive cells and rabbit tissues were examined. In addition, the nutritional and chemical composition of both bee products was investigated. Fresh and freeze-dried samples were used to isolate probiotic bacteria identified by MALDI-TOF MS Biotyper. Lactobacilli, especially <em>Apilactobacillus kunkeii, Lactiplantibacillus pentosus, Lactiplantibacillus plantarum,</em> and <em>Secundilactobacillus malefermentas</em>, received the highest identification scores. The best results were observed when targeting <em>S. aureus, S. epidermidis</em>, and <em>S. vitulinus</em>. The freeze-dried materials were tested on rabbit spermatozoa and testicular fragments to ascertain the effects of DM and DL. The results showed that the samples significantly reduced the amount of reactive oxygen species in the reproductive structures which resulted in a decrease in lipid peroxidation. Furthermore, testicular fragments treated <em>in vitro</em> with DM and DL produced considerably more testosterone and cholesterol; crude protein, nitrogenous compounds and fat were all present in higher amounts in DL while the content of reducing sugars was higher in DM.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"149 ","pages":"Pages 144-157"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143139496","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.procbio.2024.12.003
Zhaofeng Guan , Lina Zhou , Hao Wu , Zhenyu Chu , Mingjun Liu , Jun Zhou
In this study, ginsenosides and polysaccharides were extracted from ginseng residue (GR) using a green method combining deep eutectic solvent (DES) and ultrasound. The results showed that the optimal extraction conditions of ginsenosides optimized by orthogonal test were choline chloride/propionic acid ratio of 1:3, moisture content of 30 %, solid-liquid ratio of 1:25, and ultrasound time of 30 min, which resulted in the yield of ginsenosides of 36.76 ± 1.46 mg/g, and improved the efficiency by 218.27 % compared with that of the traditional 70 % ethanol extraction method. The ginseng polysaccharides obtained by further purification of the extract showed different in vitro antioxidant activities, with scavenging rates of 91.2 %, 98.3 %, and 96.4 % for hydroxyl radicals, DPPH, and superoxide radicals, respectively, and Fe(Ⅱ) chelating activity at 93.5 %. The extracted GR showed good adsorption properties for glucose, sodium nitrite, cholesterol, and sodium cholate by in vitro adsorption tests with adsorption rates of 60 %, 93.3 %, 87.05 %, and 72.95 %, respectively. In conclusion, the use of DES as an extraction solvent has certain application prospects for GR reuse.
{"title":"Deep-eutectic solvent extraction of ginsenosides and ginseng polysaccharides from ginseng residue and utilization of post-extraction residue","authors":"Zhaofeng Guan , Lina Zhou , Hao Wu , Zhenyu Chu , Mingjun Liu , Jun Zhou","doi":"10.1016/j.procbio.2024.12.003","DOIUrl":"10.1016/j.procbio.2024.12.003","url":null,"abstract":"<div><div>In this study, ginsenosides and polysaccharides were extracted from ginseng residue (GR) using a green method combining deep eutectic solvent (DES) and ultrasound. The results showed that the optimal extraction conditions of ginsenosides optimized by orthogonal test were choline chloride/propionic acid ratio of 1:3, moisture content of 30 %, solid-liquid ratio of 1:25, and ultrasound time of 30 min, which resulted in the yield of ginsenosides of 36.76 ± 1.46 mg/g, and improved the efficiency by 218.27 % compared with that of the traditional 70 % ethanol extraction method. The ginseng polysaccharides obtained by further purification of the extract showed different in vitro antioxidant activities, with scavenging rates of 91.2 %, 98.3 %, and 96.4 % for hydroxyl radicals, DPPH, and superoxide radicals, respectively, and Fe(Ⅱ) chelating activity at 93.5 %. The extracted GR showed good adsorption properties for glucose, sodium nitrite, cholesterol, and sodium cholate by in vitro adsorption tests with adsorption rates of 60 %, 93.3 %, 87.05 %, and 72.95 %, respectively. In conclusion, the use of DES as an extraction solvent has certain application prospects for GR reuse.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"149 ","pages":"Pages 99-110"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143139499","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The zein-quercetin covalent complexes were prepared by alkali treatment (A-ZEIN-Q) and free radical grafting (R-ZEIN-Q) and subsequently followed by encapsulation of oregano essential oil (OEO), and alkali grafted zein-quercetin covalent nanoparticles (A-ZQNPs-OEO) and free radical grafted zein-quercetin covalent nanoparticles (R-ZQNPs-OEO) were synthesized respectively. Additionally, the particle size, ζ-potential, microstructure, encapsulation efficiency, release behaviour, surface hydrophobicity, stability, antioxidant activity, and antibacterial activity of these nanoparticles were characterized. The results of infrared spectroscopy showed that a covalent bond was formed between zein and quercetin, the particle size of the nanoparticles decreased, and the degree of densification was higher than that of zein nanoparticles. Unlike the aggregated zein nanoparticles, the zein-quercetin nanoparticles were dispersed more uniformly. In addition, the encapsulation efficiency of OEO in A-ZQNPs-OEO and R-ZQNPs-OEO increased significantly from 45.11 % to 73.42 % and 70.90 % to that of OEO in single zein nanoparticles, respectively. Among the four nanoparticles, A-ZQNPs-OEO shows the highest stability, while the OEO-encapsulated quercetin covalent nanoparticles show better-sustained release performance, lower surface hydrophobicity, and excellent antioxidant and antibacterial activities. Overall, the synthesized OEO encapsulated zein-quercetin covalent nanoparticles possess the ability to be applied in food preservation and other applications.
{"title":"Zein-quercetin covalent nanoparticles encapsulating oregano essential oil: Improved stability, antioxidant, and antibacterial properties","authors":"Lingyu Yin, Yuhang Cao, Meihui Wang, Baohua Kong, Qian Liu, Hui Wang, Hao Wang","doi":"10.1016/j.procbio.2024.12.015","DOIUrl":"10.1016/j.procbio.2024.12.015","url":null,"abstract":"<div><div>The zein-quercetin covalent complexes were prepared by alkali treatment (A-ZEIN-Q) and free radical grafting (R-ZEIN-Q) and subsequently followed by encapsulation of oregano essential oil (OEO), and alkali grafted zein-quercetin covalent nanoparticles (A-ZQNPs-OEO) and free radical grafted zein-quercetin covalent nanoparticles (R-ZQNPs-OEO) were synthesized respectively. Additionally, the particle size, ζ-potential, microstructure, encapsulation efficiency, release behaviour, surface hydrophobicity, stability, antioxidant activity, and antibacterial activity of these nanoparticles were characterized. The results of infrared spectroscopy showed that a covalent bond was formed between zein and quercetin, the particle size of the nanoparticles decreased, and the degree of densification was higher than that of zein nanoparticles. Unlike the aggregated zein nanoparticles, the zein-quercetin nanoparticles were dispersed more uniformly. In addition, the encapsulation efficiency of OEO in A-ZQNPs-OEO and R-ZQNPs-OEO increased significantly from 45.11 % to 73.42 % and 70.90 % to that of OEO in single zein nanoparticles, respectively. Among the four nanoparticles, A-ZQNPs-OEO shows the highest stability, while the OEO-encapsulated quercetin covalent nanoparticles show better-sustained release performance, lower surface hydrophobicity, and excellent antioxidant and antibacterial activities. Overall, the synthesized OEO encapsulated zein-quercetin covalent nanoparticles possess the ability to be applied in food preservation and other applications.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"149 ","pages":"Pages 248-259"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143139492","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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