{"title":"Synergistic protease-lipase treatment for enhanced blood stain removal from textiles: Process optimization and efficacy evaluation","authors":"Mahammed Ilyas Khazi , Dilek Kut , Fakhra Liaqat , Elif Demirkan","doi":"10.1016/j.procbio.2025.05.009","DOIUrl":"10.1016/j.procbio.2025.05.009","url":null,"abstract":"<div><div>This study investigates the synergistic effects of protease and lipase, produced from strains of <em>Bacillus subtilis</em> and <em>Bacillus cereus</em>, respectively, on blood stain removal from polyester and cotton fabrics. The effectiveness of enzyme - detergent combinations was evaluated using ΔE values as a quantitative measure of stain removal. Optimal culture conditions for enzyme production were determined using Response Surface Methodology (RSM) with Central Composite Design (CCD), identifying key variables such as temperature, pH, agitation, inoculum volume, and incubation time. Under optimized conditions, the enzymatic activity of protease and lipase reached 925.5 U/ml and 20.5 U/ml, respectively, representing a 2.4-fold and 3.1-fold increase compared to unoptimized conditions (376 U/ml for protease and 6.6 U/ml for lipase). Under optimal destaining conditions (pH 7.0, enzyme concentration of 1 mg/ml, and temperature of 60 °C), the combination of detergent, protease, and lipase achieved the highest ΔE values (58.42 ± 0.5 for polyester and 6.60 ± 0.5 for cotton fabric), signifying enhanced stain removal efficacy. Synergy between enzymes and detergents was highlighted, where detergent improved enzyme access to stain components, resulting in markedly improved cleaning efficiency compared to treatments with either enzyme or detergent alone. This study emphasizes the efficacy of enzyme-assisted formulations for sustainable textile cleaning applications.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"156 ","pages":"Pages 20-32"},"PeriodicalIF":3.7,"publicationDate":"2025-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144116594","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}
Yiyang Tan , Yanhua Zhang , Jia Su , Hui Yu , Depei Wang , Xianli Xue
{"title":"Improving citric acid production in Aspergillus niger by overexpression of citrate synthase with strong promoter","authors":"Yiyang Tan , Yanhua Zhang , Jia Su , Hui Yu , Depei Wang , Xianli Xue","doi":"10.1016/j.procbio.2025.05.005","DOIUrl":"10.1016/j.procbio.2025.05.005","url":null,"abstract":"<div><div><em>Aspergillus niger</em> CGMCC10142, a strain widely used for industrial citric acid (CA) production, encodes four citrate synthase genes (<em>cs1-cs4</em>), yet their specific roles in CA biosynthesis remain poorly understood. We found that the transcript levels of the genes encoding mitochondrial citrate synthases, <em>cs3</em> or <em>cs4</em> were significantly higher than those encoding cytoplasmic citrate synthases <em>cs1</em> and <em>cs2</em> with 94.26–766.0 folds or 219.33–1782.25 folds, from 27.8 (<em>cs2</em>) to 49546.6 (<em>cs4</em>) at 24 h during CA fermentation. Given the naturally low transcription levels and cytoplasmic localization of <em>cs1</em> and <em>cs2</em>, we hypothesized that their overexpression would enhance CA production by increasing the CA available for export. Transformants overexpressing cytoplasmic <em>cs1</em> (T-c1–19) and <em>cs2</em> (T-c2–81) showed increases in CA production of 5.38 % and 18.39 %, respectively. In contrast, no significant change was observed in transformant overexpressing mitochondrial <em>cs3</em> (T-c3–11). The results showed a 6.8 % increase in acid production by strain T-c2–81, reaching 181.2 g/L at 56 h in 30 L bioreactors, compared to the starting strain of 169.6.g/L. qRT-PCR analysis of revealed that the relative expression levels of <em>cs2</em> in the T-c2–81 strain was 8.6 times higher than in the wild-type strain at 24 h. In addition, the expression levels of key genes <em>pfk</em>, <em>pc</em> and the citrate transporter <em>cexA</em> were significantly increased, ranging from 1.3 to 67 times than the control levels. This 6.8 % improvement of CA yield not only reduces unit fermentation costs but also represents a significant breakthrough in regulating intracellular metabolic flow.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"156 ","pages":"Pages 33-46"},"PeriodicalIF":3.7,"publicationDate":"2025-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144124446","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}
Nayara K.B. Lino , Gabriel L. Arruda , Felipe R. Lourenço , Valéria C. Santos-Ebinuma , Silvio S. da Silva , Júlio C. Santos , André M. Lopes
{"title":"Designing salt-induced polymer-polymer biphasic systems for optimized extraction of Monascus ruber biopigments obtained by fermentation of xylose-based medium","authors":"Nayara K.B. Lino , Gabriel L. Arruda , Felipe R. Lourenço , Valéria C. Santos-Ebinuma , Silvio S. da Silva , Júlio C. Santos , André M. Lopes","doi":"10.1016/j.procbio.2025.05.004","DOIUrl":"10.1016/j.procbio.2025.05.004","url":null,"abstract":"<div><div>The widespread use of synthetic colorants raises biosafety and environmental concerns, highlighting the need for natural alternatives like <em>Monascus</em> biopigments. These pigments possess antimicrobial and antioxidant properties, offering biotechnological and biopharmaceutical potential. Cost-effective biopigment production using xylose-rich hydrolysates from lignocellulosic biomass has been explored, but its sustainable downstream processing is crucial. Traditional liquid-liquid extraction with volatile organic compounds (VOCs) is harmful, prompting the use of eco-friendly aqueous biphasic systems (ABS) based on green chemistry principles. This study investigated the stability, partitioning, and purification of <em>Monascus ruber</em> biopigments (yellow, orange, and red) from fermented broth using ABS composed of polyethylene glycol (PEG), sodium polyacrylate (NaPA), and salts (MgSO<sub>4</sub> and CaCl<sub>2</sub>). Binodal curves showed MgSO<sub>4</sub> enhanced phase separation more effectively than CaCl<sub>2</sub>. Biopigments demonstrated high stability over 65 days and preferentially partitioned into the PEG-rich phase (<em>K</em><sub>Biopigments</sub> = 3–14), with recovery (<em>REC</em><sub>Top</sub>) values of 52–79 %. Optimal ABS conditions (18 % wt/wt PEG8000, 10 % wt/wt NaPA1200, and 0.3 M MgSO<sub>4</sub>) yielded biopigment <em>REC</em><sub>Top</sub> of 71–80 %, <em>K</em><sub>Biopigments</sub> of 12–15, and high selectivity over proteins (<em>S</em> = 14–19). Protein contaminants predominantly partitioned into the NaPA-rich phase, achieving a 10-fold purification. These findings support the stabilization and eco-friendly extraction of biopigments for bio-industrial applications.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"156 ","pages":"Pages 8-19"},"PeriodicalIF":3.7,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144105630","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}
{"title":"Continuous production of tagatose through isomerization of galactose using a tubular reactor filled with eggshell powder","authors":"Shin Futamata, Yuichiro Onishi, Fumito Tani, Takashi Kobayashi","doi":"10.1016/j.procbio.2025.05.008","DOIUrl":"10.1016/j.procbio.2025.05.008","url":null,"abstract":"<div><div>Galactose solution was passed through a tubular reactor filled with eggshell powder, and isomerization of galactose to rare sugars was continuously performed under subcritical conditions at 120–150°C with the reaction time (RT) between 3–37 min. Isomerization produced rare sugars (tagatose, talose, and sorbose). At 135°C and RT= 6 min, the maximum yield of tagatose (14.5 %) was achieved. Compared to this result, the yield of tagatose was lower at 150°C or 120°C, and the yield of byproducts was also especially lower at 120°C. However, when RT was prolonged to 37 min at 120°C, the yield of tagatose improved to about 10 %, while the formation of byproducts was kept under control. Furthermore, during the continuous use of the reactor (up to 30 h), the yield of byproducts significantly decreased with increasing usage time. Therefore, it was suggested that continuous contact of eggshell with the reaction mixture may change properties of the powder and affect the isomerization behavior of galactose. Although the eggshell powder became light brown after use for the reaction, an apparent change of the surface could not be observed by scanning electron microscope.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"155 ","pages":"Pages 128-133"},"PeriodicalIF":3.7,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144090344","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}
Thomas Conlon , Helen Herbert , Eva Campion , Nicolas Touzet
{"title":"Bioactivity of autotrophic and mixotrophic Tetraselmis chuii (CCAP 66/21B) extracts against H2O2-mediated stress in A549 cells.","authors":"Thomas Conlon , Helen Herbert , Eva Campion , Nicolas Touzet","doi":"10.1016/j.procbio.2025.05.007","DOIUrl":"10.1016/j.procbio.2025.05.007","url":null,"abstract":"<div><div>The trophic regime experienced by microalgae can influence their accumulation of antioxidant products such as phenolic compounds. This study investigated the antioxidant capacity and bioactivity of Dulbecco's phosphate-buffered saline (DPBS) extracts from the marine chlorophyte <em>Tetraselmis chuii</em> (CCAP 66/21B) cultivated under autotrophic and mixotrophic regimes. Compared to autotrophic conditions, mixotrophic cultivation significantly increased biomass yield (6.0-fold), soluble protein yield (5.7-fold) and Trolox-equivalent antioxidant capacity as measured by Folin-Ciocalteu (6.8-fold), DPPH (42.2-fold) and ABTS (15.3-fold) assays. For bioactivity, DPBS extracts (normalised by soluble protein content) were examined for their protective effects against hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>)-mediated apoptosis in pulmonary carcinoma epithelial cells (A549). Both autotrophic and mixotrophic-derived extracts exhibited a dose-dependent response in H<sub>2</sub>O<sub>2</sub>-stressed A549 cells, with significant cytoprotection observed at higher concentrations (200 µg soluble protein mL<sup>−1</sup>), while lower concentrations were insufficient to mitigate oxidative damage and were associated with reduced cell viability. These effects involved modulation of apoptosis-related mRNA expression (<em>BCL2</em> and <em>BAX</em>). The bioactive content of both autotrophic and mixotrophic-derived extracts exhibited similar biological activity with properties that may be beneficial in pulmonary pharmacology. Importantly, mixotrophy significantly upregulated the bioactive content of <em>T. chuii</em> (CCAP 66/21B) compared to obligate autotrophy (34.2-fold accounting for soluble protein yield and biomass generation).</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"155 ","pages":"Pages 117-127"},"PeriodicalIF":3.7,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144084382","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}
Roselin Neihsial, Waniabha J. Lyngdoh, Imtiwala, Arvind Kumar Singh
{"title":"Corrigendum to 'Biosorption isotherm and composition of Amido black 10B decolorizing metal and salt tolerant bacterial consortium' [Process Biochem. 155 (2025) 60–69]","authors":"Roselin Neihsial, Waniabha J. Lyngdoh, Imtiwala, Arvind Kumar Singh","doi":"10.1016/j.procbio.2025.05.002","DOIUrl":"10.1016/j.procbio.2025.05.002","url":null,"abstract":"","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"155 ","pages":"Page 92"},"PeriodicalIF":3.7,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143943423","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}
Luis Fernando Flores , Carlos S. Osorio-Gonzalez , Rahul Saini , Satinder Kaur Brar
{"title":"Streamlined ultrasonication-assisted lipid extraction and direct transesterification of Rhodosporidium toruloides 1588 single-cell oil","authors":"Luis Fernando Flores , Carlos S. Osorio-Gonzalez , Rahul Saini , Satinder Kaur Brar","doi":"10.1016/j.procbio.2025.05.006","DOIUrl":"10.1016/j.procbio.2025.05.006","url":null,"abstract":"<div><div><em>Rhodosporidium toruloides</em>-1588 efficiently utilizes various renewable residues for lipid synthesis, that potentially can be used for advanced drop-in biofuel production. However, to improve the feasibility, a streamlined sequential lipid extraction-transesterification process with minimal unit operations is essential. To enhance lipid extraction from <em>R. toruloides</em>-1588 fresh biomass, environmentally friendly solvents in combination with ultrasound-assisted extraction (UAE) were evaluated and compared to the highly used Folch-modified method. The results demonstrated a 291.45 % extraction efficiency in comparison to the reference method using methanol-UAE at the 1:1 ratio (broth removed: solvent added). Subsequent one- or two-step transesterification using KOH and H<sub>2</sub>SO<sub>4</sub> produced fatty acid methyl esters (FAME’s) with a similar distribution to the reference method being oleic, linoleic, palmitic, and stearic fatty acids, the most abundant. Thus, a potentially energy-saving method was developed by removing cell drying and thermochemical hydrolysis steps. Likewise, the proposed method decreases solvent consumption during the ultrasonication-assisted lipid extraction and direct transesterification of <em>R. toruloides</em>-1588 single-cell oil.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"157 ","pages":"Pages 1-9"},"PeriodicalIF":3.7,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144194940","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}
{"title":"Multi-objective optimization of industrial batch Balhimycin antibiotic and fed-batch Lysine biochemical processes","authors":"Swaprabha P. Patel , Ashish M. Gujarathi","doi":"10.1016/j.procbio.2025.05.003","DOIUrl":"10.1016/j.procbio.2025.05.003","url":null,"abstract":"<div><div>Lysine is an essential amino acid, and its market value is USD 6.8 billion in 2023, and it is expected to double by 2033 worldwide. Balhimycin belongs to an important group of antibiotics with highly complex structures. With the utilization of advanced computational knowledge and new technologies, even a small improvement in productivity, yield, process time, or product quality can bring noticeable improvement in the production technology of biochemical processes. This study proposes a jumping gene adaptation of the elitist multi-objective differential evolution (Elitist MODE) algorithm. It is tested and applied successfully on the five benchmark test problems and two real-world problems, batch Balhimycin and fed-batch Lysine fermentation. Different constraint-handling techniques are used to test the performance of the proposed algorithm. The result obtained shows that the proposed algorithm can give a widespread Pareto front for both the test and industrial optimization problems. In batch Balhimycin fermentation, the optimal amount of glycerol required is not exceeding 59.84 g/L, and the optimal uptake of phosphate is also very low and doesn’t exceed 0.5 g/L, upon which product inhibition occurs. The uncertainty-specific study reveals that, with variation in upper bound of AMS concentration, product concentration varies significantly from 0.61 to 0.75 g/L.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"155 ","pages":"Pages 93-116"},"PeriodicalIF":3.7,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144070740","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}
Shiying Li , Fang Li , Peipei Li , Yanjun Yue , Jinfang Tan , Xiaolei Jie , Wei Zhou , Yanlai Han
{"title":"Evaluation of the structure and properties of humic acids from Yunnan lignite transformed by Penicillium ortum MJ51","authors":"Shiying Li , Fang Li , Peipei Li , Yanjun Yue , Jinfang Tan , Xiaolei Jie , Wei Zhou , Yanlai Han","doi":"10.1016/j.procbio.2025.04.024","DOIUrl":"10.1016/j.procbio.2025.04.024","url":null,"abstract":"<div><div>Biotransformation of low-rank coal into humic acids (HAs) has received increasing attention. In this study, <em>Penicillium ortum</em> MJ51, isolated from coal, was used to transform Yunnan lignite into HAs. Differences in the yield, structure and properties were evaluated for HAs produced from the biodegradation of nitric acid pretreated-lignite (MHA), and those produced via chemical extraction from raw lignite (CHA<sub>1</sub>) and nitric acid pretreated-lignite (CHA<sub>2</sub>). The results show that with the progression of degradation time, the MHA yield gradually increased and peaked on day 12 at 62.39 %, which was 199.60 % and 40.68 % higher than that of CHA<sub>1</sub> and CHA<sub>2</sub>, respectively. Meanwhile, the contents of macro-micro nutrients and active groups (including carbonyl, carboxyl, methoxy, and N-alkyl), as well as the anti-flocculation ability and aliphaticity of MHA also gradually increased. However, the contents of C, H, N, heavy metals and ash, particle size, relative molecular weight, aromaticity, and hydrophobicity of MHA gradually decreased. On days 12–16 of degradation, the MHA had lower heavy metals, rare earth elements and ash content, higher macro-micro nutrients content, lower relative molecular weight and aromaticity, higher anti-flocculation ability and bioactivity than those extracted via the chemical method, making it suitable for agricultural applications.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"155 ","pages":"Pages 70-83"},"PeriodicalIF":3.7,"publicationDate":"2025-05-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143921980","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}
Yangdan Fang , Lichao Lu , Guangquan Ou , Qinqin Zhu , Zhuqiu Sun , Ziyu Liu , Marvin Yeung , Jinying Xi
{"title":"Stable biofilm formation via nutrition regulation for enhancing toluene removal in gas–solid fluidized-bed bioreactors","authors":"Yangdan Fang , Lichao Lu , Guangquan Ou , Qinqin Zhu , Zhuqiu Sun , Ziyu Liu , Marvin Yeung , Jinying Xi","doi":"10.1016/j.procbio.2025.05.001","DOIUrl":"10.1016/j.procbio.2025.05.001","url":null,"abstract":"<div><div>Gas-solid fluidized-bed bioreactors (FBRs), an innovative equipment for gaseous volatile organic compound (VOC) treatment, are used to realize efficient removal of VOCs characterized by large volumes and low concentrations and avoid clogging due to excessive biofilm accumulation. However, difficulties in biomass accumulation caused by stressful environmental pressures, especially shear force, limit their development. In this study, two laboratory-scale FBRs were built, and toluene removal was nearly twiced with saturated relative humidity and adequate nutrition supplement. Accelerated biofilm formation, activated microbial metabolism, enhanced mechanical strength, and surface smoothness were also observed. Nutrition regulation optimized cell synthesis and extracellular polymeric substance secretion through alternation of the accumulation of elements, especially C/N and Ca, which increased by 84 % and 112 %, respectively. The biofilm loss was reduced by 50–75 % under compression and collision conditions, indicating the promoted biofilm stability under shear force caused by extrusion and collision between particles. With the initial drawbacks partially mitigated, nutrient regulation during FBR operation was found to be useful. This study provides theoretical support for further applications of FBRs and efficient treatment of gaseous VOCs with large gas volumes and low concentrations.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"155 ","pages":"Pages 84-91"},"PeriodicalIF":3.7,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143921981","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}