Applied Biochemistry and Biotechnology最新文献

{"title":"Design of Bionanomaterial of Chitosan Carbohydrate Polymer Composited with Broccoli Extract and Zinc Oxide Nanoparticles: Anticancer Activity in Human Osteosarcoma.","authors":"Muthanna O Hussein, Ahmed Saud Abdulhameed","doi":"10.1007/s12010-024-05066-3","DOIUrl":"https://doi.org/10.1007/s12010-024-05066-3","url":null,"abstract":"<p><p>In the current research, a chitosan/broccoli extract/ZnO nanoparticle (CH/BE/ZnO) bionanocomposite was created. The physicochemical properties of CH/BE/ZnO bionanocomposite were investigated using a variety of methods, including field emission scanning electron microscopy (FESEM), elemental analysis (CHN-O), X-ray diffraction (XRD), Fourier transform infrared spectrum (FTIR), Brunauer-Emmett-Teller (BET), and transmission electron microscopy (TEM). The CH/BE/ZnO bionanocomposite's biological activity was assessed by examining its cytotoxicity capabilities against a bone cancer cell line (MG63). The total pore volume and specific surface area of CH/BE/ZnO are 0.134 cm³/g and 16.99 m²/g, respectively. The IC50 results for CH/BE/ZnO bionanocomposite in bone cancer investigations using the MTT test against the MG63 cell line was 115 μg/mL. The results indicate that the CH/BE/ZnO bionanocomposite is an effective chemotherapeutic agent against human osteosarcoma. The CH/BE/ZnO bionanocomposite showed high performance and structure, which means innovating nanomaterial agents for biological applications in the future.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142338715","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Antimicrobial Effects of Orthodontic Molar Tube Coated with ZnO Nanoparticles Using Electrophoretic Deposition Method: A Randomized Clinical Trial.","authors":"Ahmed K Al-Murshady, Dheaa H Al-Groosh","doi":"10.1007/s12010-024-05062-7","DOIUrl":"https://doi.org/10.1007/s12010-024-05062-7","url":null,"abstract":"<p><p>This study aimed to evaluate the antimicrobial effect of coated orthodontic molar tubes (COMT) with zinc oxide nanoparticles (ZnO NPs) using an electrophoretic deposition method (EPD) and to evaluate the orthodontic molar tubes (OMT) bond failure rate. Seventy-two orthodontic molar tubes (OMTs) for second molars were divided into two groups 36 each; one group coated with ZnO NPs and the other control negative uncoated. The OMT was coated using the EPD method with ZnO NPs in a concentration of 10 g/l. The OMTs were randomly allocated using a split-mouth, cross-quadrant design. After 2 weeks of appliance placement, swabs were taken from the surface of the OMTs for microbial assessment against Streptococcus mutans, Lactobacillus acidophilus, and total bacterial counts; additionally, plaque and gingival indices were assessed. The patient was followed for 3 months to evaluate the bond failure rate. The COMT showed a statistically significant reduction in total bacterial accounts, S. mutans, and L. acidophilus compared to UOMT (P < 0.001). Furthermore, the plaque and gingival indices near COMT were significantly less than that of UOMT. The bond failure rate was not significant between the COMT and UOMT. The COMT with ZnO NPs has potent antibacterial activity against the tested pathogens with a reduction in the amount of plaque accumulation. The use of the EPD method was feasible without adverse effects on the orthodontic molar tubes bond failure rate.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142338713","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Values of LncRNA SNHG14 in the Differential Diagnosis and Prognosis Evaluation of Acute Ischemic Stroke.","authors":"Hongxiang Lu, Yuezhan Zhang","doi":"10.1007/s12010-024-05070-7","DOIUrl":"https://doi.org/10.1007/s12010-024-05070-7","url":null,"abstract":"<p><p>The long non-coding RNA (LncRNA) SNHG14 has been investigated for its potential in acute ischemic stroke (AIS) and transient ischemic attack (TIA) diagnosis. Thirty-two healthy people, 85 patients with AIS, and 40 patients with TIA had their blood tested to determine SNHG14 mRNA transcript levels using quantitative real-time polymerase chain reaction (qRT-PCR). A stroke's severity was measured using the Stroke Severity Scale developed by the National Institutes of Health (NIHSS). After 30 days, individuals with AIS were evaluated for progress using a modified Rankin Scale (mRS). There was no significant difference in SNHG14 LncRNA levels between TIA patients and controls, despite the huge rise in AIS incidence (p > 0.05) (all p < 0.001). Compared to those who did well on the AIS test, those who performed poorly had substantially greater levels of SNHG14 LncRNA (mRS 0-1 points) (mRS 0-2). LncRNA SNHG14 had an AUC of 0.714 (80%, 61.18%) when used to identify AIS in TIA patients, and a comparable finding was seen when predicting a poor 30-day prognosis of AIS (73%, 66.67%). There are also graphical representations of the findings. Improvements in NIHSS and mRS scores were associated with increases in SNHG14 LncRNA mRNA levels in individuals diagnosed with AIS. It is critical that we focus entirely on this decision (all p < 0.05). Analysis of the long non-coding RNA known as SNHG14 in the patient's blood can be used to diagnose AIS, rule out TIA, forecast the intensity of the disease, and evaluate the prognosis. You can accomplish everything on that list simultaneously.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142338725","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Harnessing the Energy Potential and Value-Added Products from the Treatment of Sugarcane Vinasse: Maximizing Methane Production Through Co-Digestion with Sugarcane Molasses and Enhanced Organic Loading.","authors":"Alexandre Rodrigues Ribeiro, Kauanna Uyara Devens, Franciele Pereira Camargo, Isabel Kimiko Sakamoto, Maria Bernadete Amâncio Varesche, Edson Luiz Silva","doi":"10.1007/s12010-024-05078-z","DOIUrl":"https://doi.org/10.1007/s12010-024-05078-z","url":null,"abstract":"<p><p>This study assessed the impact of organic loading rate (OLR) on methane (CH₄) production in the anaerobic co-digestion (AcoD) of sugarcane vinasse and molasses (SVM) (1:1 ratio) within a thermophilic fluidized bed reactor (AFBR). The OLR ranged from 5 to 27.5 kg COD.m^-3.d^-1, with a fixed hydraulic retention time (HRT) of 24 h. Organic matter removal varied from 56 to 84%, peaking at an OLR of 5 kg COD.m^-3.d^-1. Maximum CH₄ yield (MY) (272.6 mL CH₄.g^-1COD_rem) occurred at an OLR of 7.5 kg COD.m^-3.d^-1, while the highest CH₄ production rate (MPR) (4.0 L CH₄.L^-1.d^-1) and energy potential (E.P.) (250.5 kJ.d^-1) were observed at an OLR of 20 kg COD.m^-3.d^-1. The AFBR exhibited stability across all OLR. At 22.5 kg COD.m^-3.d^-1, a decrease in MY indicated methanogenesis imbalance and inhibitory organic compound accumulation. OLR influenced microbial populations, with Firmicutes and Thermotogota constituting 43.9% at 7.5 kg COD.m^-3.d^-1, and Firmicutes dominating (52.7%) at 27.5 kg COD.m^-3.d^-1. Methanosarcina (38.9%) and hydrogenotrophic Methanothermobacter (37.6%) were the prevalent archaea at 7.5 kg COD.m^-3.d^-1 and 27.5 kg COD.m^-3.d^-1, respectively. Therefore, this study demonstrates that the organic loading rate significantly influences the efficiency of methane production and the stability of microbial communities during the anaerobic co-digestion of sugarcane vinasse and molasses, indicating that optimized conditions can maximize energy yield and maintain methanogenic balance.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142338718","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Electro-acupuncture Suppresses Ferroptosis to Alleviate Cerebral Ischemia-Reperfusion Injury Through KAT3B-Mediated Succinylation of ACSL4.","authors":"Fang Liu, Ying Chen, Kangbai Huang","doi":"10.1007/s12010-024-05063-6","DOIUrl":"https://doi.org/10.1007/s12010-024-05063-6","url":null,"abstract":"<p><p>Electro-acupuncture (EA) is identified as an effective therapeutic method for cerebral ischemia/reperfusion injury (CIRI), which is a combination of Chinese traditional acupuncture and modern electro-therapy. However, the downstream molecular mechanisms of EA in CIRI process remains largely unknown. The purpose of the present study is to unveil the therapeutic effect of EA on CIRI rat and its regulatory mechanisms. At first, we constructed middle cerebral artery occlusion (MCAO) rat models and then treated them with EA to observe the pathological changes. The results indicated that EA decreased the infarct volume (43.81 ± 3.34 vs 15.96 ± 2.22) and the neurological scores (3.33 ± 0.52 vs 1.67 ± 0.52) and suppressed the apoptosis in MCAO model rats. For ferroptosis analysis, EA decreased the Fe2 + (0.08 ± 0.01 vs 0.06 ± 0.01), MDA (36.61 ± 4.29 vs 21.72 ± 2.79), and LPS (5.25 ± 0.69 vs 2.89 ± 0.42) contents and increased the GSH (4.94 ± 1.04 vs 11.69 ± 1.88) content in MCAO model rats. We next detected whether succinylation mediated EA-treated I/R injury. According to immunoprecipitation and western blot analysis, EA treatment could lower both levels of succinylation and KAT3B in MCAO rats. Moreover, mechanism experiments unveiled that KAT3B promoted the succinylation of the ferroptosis-related protein ACSL4 at K661 site and thus stabilizing ACSL4. Finally, EA-treated MCAO rats were further injected with KAT3B expression vector. The results showed that KAT3B overexpression increased the infarct volume (31.44 ± 3.92 vs 7.94 ± 2.84) and the neurological scores (2.67 ± 0.51 vs 1.33 ± 0.51) and promoted the apoptosis in EA treated MCAO model rats. For ferroptosis analysis, KAT3B overexpression increased the Fe2 + (0.08 ± 0.01 vs 0.05 ± 0.01), MDA (29.24 ± 4.30 vs 22.06 ± 1.89), and LPO (5.07 ± 0.45 vs 2.88 ± 0.49) contents and decreased the GSH (7.86 ± 1.09 vs 11.06 ± 1.76) content in EA treated MCAO model rats. Collectively, our study demonstrates that EA plays a therapeutic role in CIRI through suppressing KAT3B-induced stabilization of ACSL4 to inhibit ferroptosis. These findings contribute to our understanding of the molecular mechanisms underlying the neuroprotective effects of EA and open new avenues for the development of innovative therapeutic strategies for CIRI.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142338717","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of Yeast Strain YA176 for Bio-Purification of Soy Molasses to Produce Raffinose Family Oligosaccharides and Optimization of Fermentation Conditions.","authors":"Zhilei Fu, Shuang Cheng, Jinghao Ma, Rana Abdul Basit, Yihua Du, Shubin Tian, Guangsen Fan","doi":"10.1007/s12010-024-05065-4","DOIUrl":"https://doi.org/10.1007/s12010-024-05065-4","url":null,"abstract":"<p><p>Soybean molasses, which contains high levels of raffinose family oligosaccharides (RFOs) such as stachyose and raffinose, is subjected to a process of bio-purification to remove sucrose while maintaining the RFOs, consequently increasing its value. This study employed morphological observation, physiological and biochemical studies, and molecular biology techniques to identify YA176, a yeast strain renowned for its effective bio-purification of soy molasses. Through single-factor and orthogonal experiments, optimal bio-purification conditions were established. YA176, belonging to Wickerhamomyces anomalus, demonstrated robust growth across a wide range of temperature and pH levels, coupled with remarkable tolerance to glucose, sucrose, and NaCl up to 41.2%, 47.3%, and 10%, respectively. Under these optimized conditions, YA176 efficiently utilized sucrose while preserving 93.3% of raffinose and 78.6% of stachyose, ensuring the retention of functional RFOs. In summary, yeast strain YA176 exhibits exceptional bio-purification abilities, making it an ideal candidate for producing functional RFOs from soy molasses.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142338720","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Production, Characterization, Kinetics, and Thermodynamics Analysis of Amyloglucosidase from Fungal Consortium.","authors":"Imran Ali, Roheena Abdullah, Asifa Saleem, Kinza Nisar, Afshan Kaleem, Mehwish Iqtedar, Irfana Iqbal, Xiaoming Chen","doi":"10.1007/s12010-024-05067-2","DOIUrl":"https://doi.org/10.1007/s12010-024-05067-2","url":null,"abstract":"<p><p>The current study aimed to produce an amyloglucosidase enzyme from the fungal consortium. The best amylolytic fungal consortia were identified as Alternaria alternata and Aspergillus niger through the 18S rDNA technique. Fermentation kinetics and various nutritional and cultural parameters were analyzed. Maximum production was obtained in M4 media, pH 5.5, 30 °C, and 4 mL inoculum at 150 rpm after 72 h of incubation. Along with that, sodium nitrate at 2.5%, maltose, beef extract 1%, zinc sulfate (0.1%), and Tween 80 (0.1%) supported the maximum amyloglucosidase production. Amyloglucosidase was partially purified up to 1.6 purification fold with a specific activity of 1.84 Umg^-1 in a stepwise manner by ammonium sulfate purification, dialysis, and ion exchange chromatography. The AMG enzyme also revealed maximum activity at 50 °C with 5.0 pH. Upon the kinetic analysis, the specific yield coefficient Yp/x and volumetric rates Qp and Qx were also found to be significant in the above optimized conditions. The Km value 0.33 mg mL^-1 and Vmax 26.31 U mL^-1 were obtained at 1% soluble starch substrate. Thermodynamic parameters for soluble starch hydrolysis were as follows: ΔH = 48.78 kJ mol^-1, (Ea) =  - 46.0 kJ mol^-1, and ΔS =  - 43.10 J mol^-1 K^-1. This finding indicates the indigenously isolated fungal consortium can be the best candidate for industrial applications.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142338724","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High-Level Expression of Sucrose Isomerase in Bacillus subtilis Through Expression Element Optimization and Fermentation Optimization.","authors":"Kang Zhang, Wenchong Zhao, Sheng Chen, Lingqia Su, Jing Wu","doi":"10.1007/s12010-024-05042-x","DOIUrl":"https://doi.org/10.1007/s12010-024-05042-x","url":null,"abstract":"<p><p>Sucrose isomerase is an important food enzyme that catalyzes the isomerization of sucrose into isomaltulose, a functional sugar widely used in food industry, while the production level of sucrose isomerase in food safe host strains was much lower than industrial requirement. Bacillus subtilis is an excellent host strain for recombinant protein expression, which owns the characteristics of powerful secretory capability and generally recognized as safe state. In this study, the expression of sucrose isomerase in B. subtilis was improved through expression element optimization and fermentation optimization. Firstly, the extracellular chaperone PrsA was overexpressed to enhance extracellular folding of sucrose isomerase, which improved the recombinant expression level by 80.02%. Then, the protein synthesis level was optimized through promoter screening, improving the recombinant expression level by 60.40%. On the basis of strain modification, the fermentation conditions including nitrogen source, carbon source, metal ion, pH and temperature were optimized successively in shake-flask. Finally, the 3 L bioreactor cultivation condition was optimized and yielding a sucrose isomerase activity of 862.86 U/mL, the highest level among the food safety strains. This study provides an effective strategy to improve the expression level of food enzymes in B. subtilis.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142338719","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Self-Powered Enzymatic Glucose Sensor Utilizing Bimetallic Nanoparticle Composites Modified Pencil Graphite Electrodes as Cathode.","authors":"Gamze Emir, Yusuf Dilgin, Samet Şahin, Cahit Akgul","doi":"10.1007/s12010-024-05068-1","DOIUrl":"https://doi.org/10.1007/s12010-024-05068-1","url":null,"abstract":"<p><p>Enzymatic biofuel cells (EBFC) are promising sources of green energy owing to the benefits of using renewable biofuels, eco-friendly biocatalysts, and moderate operating conditions. In this study, a simple and effective EBFC was presented using an enzymatic composite material-based anode and a nonenzymatic bimetallic nanoparticle-based cathode respectively. The anode was constructed from a glassy carbon electrode (GCE) modified with a multi-walled carbon nanotube (MWCNT) and ferrocene (Fc) as a conductive layer coupled with the enzyme glucose oxidase (GOx) as a sensitive detection layer for glucose. A chitosan layer was also applied to the electrode as a protective layer to complete the composite anode. Chronoamperometry (CA) results show that the MWCNT-Fc-GOx/GCE electrode has a linear relationship between current and glucose concentration, which varied from 1 to 10 mM. The LOD and LOQ were calculated for anode as 0.26 mM and 0.87 mM glucose, respectively. Also the sensitivity of the proposed sensor was calculated as 25.71 <math><mi>μ</mi></math> A/mM. Moreover, the studies of some potential interferants show that there is no significant interference for anode in the determination of glucose except ascorbic acid (AA), uric acid (UA), and dopamine (DA). On the other hand, the cathode consisted of a disposable pencil graphite electrode (PGE) modified with platinum-palladium bimetallic nanoparticles (Nps) which exhibit excellent conductivity and electron transfer rate for the oxygen reduction reaction (ORR). The constructed EBFC was optimized and characterized using various electroanalytical techniques. The EBFC consisting of MWCNT-Fc-GOx/GCE anode and Pt-PdNps/PGE cathode exhibits an open circuit potential of 285.0 mV and a maximum power density of 32.25 µW cm^-2 under optimized conditions. The results show that the proposed EBFC consisting of an enzymatic composite-based anode and bimetallic nanozyme-based cathode is a unique design and a promising candidate for detecting glucose while harvesting power from glucose-containing natural or artificial fluids.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142338704","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Construction of an Aspergillus oryzae △nptB△pyrG Host for Homologous Expression of Lipase and Catalytic Property Characterization of Recombinant Lipase.","authors":"Yueting Zhang, Hongmei Nie, Fei Zhang, Mengmeng Jin, Zhao Wang, Jianyong Zheng","doi":"10.1007/s12010-024-05064-5","DOIUrl":"https://doi.org/10.1007/s12010-024-05064-5","url":null,"abstract":"<p><p>Aspergillus oryzae is an ideal cell factory for protein expression with powerful protein processing and secretion capabilities. The current study aimed to explore the homologous expression of A. oryzae lipase AOL (GenBank: KP975533) by constructing an auxotrophic A. oryzae △pyrG△nptB and subsequently characterizing the immobilization and catalytic properties of recombinant lipase. Initially, the pyrG gene knocked out in wild-type A. oryzae by homologous recombination, followed by the creation of a uridine/uracil auxotroph transformation. Through this system, the protease gene nptB was precisely knocked out, leading to a substantial decrease in extracellular (39.04%) and intracellular (90.07%) protease activity. The A. oryzae △nptB△pyrG strain was used as host for homologous expression of lipase AOL. After transformation of linearized lipase-expression cassette, the engineered A. oryzae AOL-8 was screened out with the lipase gene copy number of 14, exhibiting extracellular and intracellular lipase activities of 1.75 U/mL and 46.4 U/g, respectively. Subsequently, the production and immobilization of the recombinant lipase, via physical adsorption on macroporous resin XRZ04B, were achieved through submerged fermentation of the AOL-8 strain. The results of esterification catalytic properties of immobilized recombinant lipase indicated that the lipase exhibited optimal catalytic activity with lauric acid and methanol as substrates, a reaction temperature of 35 °C, and n-hexane as the preferred solvent medium; its highest conversion rate can reach at 72.3%.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142338714","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}