{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T22:25:57Z","timestamp":1782253557765,"version":"3.54.5"},"reference-count":44,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2022,7,14]],"date-time":"2022-07-14T00:00:00Z","timestamp":1657756800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["21977020"],"award-info":[{"award-number":["21977020"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2019QN01N107"],"award-info":[{"award-number":["2019QN01N107"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2020B121201009"],"award-info":[{"award-number":["2020B121201009"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2020GDASYL-20200301002"],"award-info":[{"award-number":["2020GDASYL-20200301002"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"the Department of Science and Technology of Guangdong Province","award":["21977020"],"award-info":[{"award-number":["21977020"]}]},{"name":"the Department of Science and Technology of Guangdong Province","award":["2019QN01N107"],"award-info":[{"award-number":["2019QN01N107"]}]},{"name":"the Department of Science and Technology of Guangdong Province","award":["2020B121201009"],"award-info":[{"award-number":["2020B121201009"]}]},{"name":"the Department of Science and Technology of Guangdong Province","award":["2020GDASYL-20200301002"],"award-info":[{"award-number":["2020GDASYL-20200301002"]}]},{"name":"Guangdong Provincial Key Laboratory","award":["21977020"],"award-info":[{"award-number":["21977020"]}]},{"name":"Guangdong Provincial Key Laboratory","award":["2019QN01N107"],"award-info":[{"award-number":["2019QN01N107"]}]},{"name":"Guangdong Provincial Key Laboratory","award":["2020B121201009"],"award-info":[{"award-number":["2020B121201009"]}]},{"name":"Guangdong Provincial Key Laboratory","award":["2020GDASYL-20200301002"],"award-info":[{"award-number":["2020GDASYL-20200301002"]}]},{"name":"GDAS\u2019 Project of Science and Technology Development","award":["21977020"],"award-info":[{"award-number":["21977020"]}]},{"name":"GDAS\u2019 Project of Science and Technology Development","award":["2019QN01N107"],"award-info":[{"award-number":["2019QN01N107"]}]},{"name":"GDAS\u2019 Project of Science and Technology Development","award":["2020B121201009"],"award-info":[{"award-number":["2020B121201009"]}]},{"name":"GDAS\u2019 Project of Science and Technology Development","award":["2020GDASYL-20200301002"],"award-info":[{"award-number":["2020GDASYL-20200301002"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Expression and purification of \u03b2-galactosidases derived from Bifidobacterium provide a new resource for efficient lactose hydrolysis and lactose intolerance alleviation. Here, we cloned and expressed two \u03b2-galactosidases derived from Bifidobacterium. The optimal pH for BLGLB1 was 5.5, and the optimal temperature was 45 \u00b0C, at which the enzyme activity of BLGLB1 was higher than that of commercial enzyme E (300 \u00b1 3.6 U\/mg) under its optimal conditions, reaching 2200 \u00b1 15 U\/mg. The optimal pH and temperature for BPGLB1 were 6.0 and 45 \u00b0C, respectively, and the enzyme activity (0.58 \u00b1 0.03 U\/mg) under optimum conditions was significantly lower than that of BLGLB1. The structures of the two \u03b2-galactosidase were similar, with all known key sites conserved. When o-nitrophenyl-\u03b2-D-galactoside (oNPG) was used as an enzyme reaction substrate, the maximum reaction velocity (Vmax) for BLGLB1 and BPGLB1 was 3700 \u00b1 100 U\/mg and 1.1 \u00b1 0.1 U\/mg, respectively. The kinetic constant (Km) of BLGLB1 and BPGLB1 was 1.9 \u00b1 0.1 and 1.3 \u00b1 0.3 mmol\/L, respectively. The respective catalytic constant (kcat) of BLGLB1 and BPGLB1 was 1700 \u00b1 40 s\u22121 and 0.5 \u00b1 0.02 s\u22121, respectively; the respective kcat\/Km value of BLGLB1 and BPGLB1 was 870 L\/(mmol\u2219s) and 0.36 L\/(mmol\u2219s), respectively. The Km, kcat and Vmax values of BLGLB1 were superior to those of earlier reported \u03b2-galactosidase derived from Bifidobacterium. Overall, BLGLB1 has potential application in the food industry.<\/jats:p>","DOI":"10.3390\/molecules27144497","type":"journal-article","created":{"date-parts":[[2022,7,15]],"date-time":"2022-07-15T01:57:11Z","timestamp":1657850231000},"page":"4497","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Cloning, Expression, Purification, and Characterization of \u03b2-Galactosidase from Bifidobacterium longum and Bifidobacterium pseudocatenulatum"],"prefix":"10.3390","volume":"27","author":[{"given":"Mingzhu","family":"Du","sequence":"first","affiliation":[{"name":"School of Food Science and Technology, Jiangnan University, Wuxi 214122, China"},{"name":"Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuanghong","family":"Yang","sequence":"additional","affiliation":[{"name":"School of Food Science and Technology, Jiangnan University, Wuxi 214122, China"},{"name":"Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tong","family":"Jiang","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tingting","family":"Liang","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4012-6954","authenticated-orcid":false,"given":"Ying","family":"Li","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuzhen","family":"Cai","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6503-359X","authenticated-orcid":false,"given":"Qingping","family":"Wu","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jumei","family":"Zhang","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Chen","sequence":"additional","affiliation":[{"name":"School of Food Science and Technology, Jiangnan University, Wuxi 214122, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3943-7755","authenticated-orcid":false,"given":"Xinqiang","family":"Xie","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/j.bcab.2015.12.010","article-title":"Lactose hydrolysis approach: Isolation and production of \u03b2-galactosidase from newly isolated Bacillus strain B-2","volume":"5","author":"Kamran","year":"2016","journal-title":"Biocatal. Agric. Biotechnol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1186\/s12934-015-0214-8","article-title":"Heterologous expression of a recombinant lactobacillal \u03b2-galactosidase in Lactobacillus plantarum: Effect of different parameters on the sakacin P-based expression system","volume":"14","author":"Nguyen","year":"2015","journal-title":"Microb. Cell Factories"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1016\/j.biotechadv.2011.03.008","article-title":"Recombinant microbial systems for improved \u03b2-galactosidase production and biotechnological applications","volume":"29","author":"Oliveira","year":"2011","journal-title":"Biotechnol. Adv."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Cheng, J., Romantsov, T., Engel, K., Doxey, A.C., Rose, D.R., Neufeld, J.D., and Charles, T.C. (2017). Functional metagenomics reveals novel \u03b2-galactosidases not predictable from gene sequences. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0172545"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/S1369-703X(01)00118-8","article-title":"Effects of temperature and pH on the catalytic activity of the immobilized \u03b2-galactosidase from Kluyveromyces lactis","volume":"9","author":"Zhou","year":"2001","journal-title":"Biochem. Eng. J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1016\/j.biortech.2019.01.005","article-title":"\u03b2-Galactosidase from Exiguobacterium acetylicum: Cloning, expression, purification and characterization","volume":"277","author":"Aburto","year":"2019","journal-title":"Bioresour. Technol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3074","DOI":"10.1111\/ijfs.14222","article-title":"Whey valorization using transgalactosylation activity of immobilized \u03b2-galactosidase","volume":"54","author":"Banjanac","year":"2019","journal-title":"Int. J. Food Sci. Technol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.1007\/s12010-016-2078-8","article-title":"Utilization of Cheese Whey Using Synergistic Immobilization of beta-Galactosidase and Saccharomyces cerevisiae Cells in Dual Matrices","volume":"179","author":"Kokkiligadda","year":"2016","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_9","first-page":"S31","article-title":"Lactose intolerance in Indonesian children","volume":"24","author":"Hegar","year":"2015","journal-title":"Asia Pac. J. Clin. Nutr."},{"key":"ref_10","first-page":"S1","article-title":"From \u2018lactose intolerance\u2019 to \u2018lactose nutrition\u2019","volume":"24","author":"Lukito","year":"2015","journal-title":"Asia Pac. J. Clin. Nutr."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"113","DOI":"10.2147\/CEG.S32368","article-title":"Lactose intolerance: Diagnosis, genetic, and clinical factors","volume":"5","author":"Mattar","year":"2012","journal-title":"Clin. Exp. Gastroenterol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.procbio.2019.04.016","article-title":"A novel cold-adapted \u03b2-galactosidase from Alteromonas sp. ML117 cleaves milk lactose effectively at low temperature","volume":"82","author":"Yao","year":"2019","journal-title":"Process Biochem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"108746","DOI":"10.1016\/j.lwt.2019.108746","article-title":"Improvement in the yield and selectivity of lactulose synthesis with Bacillus circulans \u03b2-galactosidase","volume":"118","author":"Aburto","year":"2020","journal-title":"LWT"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1007\/s13205-017-0645-5","article-title":"Sources of \u03b2-galactosidase and its applications in food industry","volume":"7","author":"Saqib","year":"2017","journal-title":"3 Biotech"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1016\/j.bioorg.2018.01.006","article-title":"Purification and characterization of \u03b2-galactosidase from probiotic Pediococcus acidilactici and its use in milk lactose hydrolysis and galactooligosaccharide synthesis","volume":"77","author":"Chanalia","year":"2018","journal-title":"Bioorg. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"129050","DOI":"10.1016\/j.foodchem.2021.129050","article-title":"\u03b2-galactosidase from Kluyveromyces lactis in genipin-activated chitosan: An investigation on immobilization, stability, and application in diluted UHT milk","volume":"349","author":"Lima","year":"2021","journal-title":"Food Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.nbt.2021.03.002","article-title":"A novel thermophile \u03b2-galactosidase from Thermothielavioides terrestris producing galactooligosaccharides from acid whey","volume":"63","author":"Zerva","year":"2021","journal-title":"New Biotechnol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1111\/j.1574-6968.2006.00614.x","article-title":"Characterization and molecular cloning of a heterodimeric \u03b2-galactosidase from the probiotic strain Lactobacillus acidophilus R22","volume":"269","author":"Nguyen","year":"2007","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1007\/s00253-001-0911-6","article-title":"Purification and characterization of a recombinant beta-galactosidase with transgalactosylation activity from Bifidobacterium infantis HL96","volume":"58","author":"Hung","year":"2002","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"108545","DOI":"10.1016\/j.lwt.2019.108545","article-title":"Purification, characterization and thermal inactivation kinetics of \u03b2-galactosidase from Lactobacillus leichmannii 313","volume":"116","author":"Ji","year":"2019","journal-title":"LWT"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1717","DOI":"10.4014\/jmb.1905.05068","article-title":"Cloning and Heterologous Expression of the \u03b2-Galactosidase Gene from Bifidobacterium longum RD47 in B. bifidum BGN4","volume":"29","author":"Park","year":"2019","journal-title":"J. Microbiol. Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1007\/s00217-013-1938-5","article-title":"Lactobacillus plantarum 70810 from Chinese paocai as a potential source of \u03b2-galactosidase for prebiotic galactooligosaccharides synthesis","volume":"236","author":"Zhang","year":"2013","journal-title":"Eur. Food Res. Technol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1007\/BF02817446","article-title":"Isolation and characterization of \u03b2-galactosidase fromLactobacillus crispatus","volume":"45","author":"Kim","year":"2000","journal-title":"Folia Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1007\/s12011-009-8519-2","article-title":"Enhancement of \u03b1- and \u03b2-Galactosidase Activity in Lactobacillus reuteri by Different Metal Ions","volume":"136","author":"Ibrahim","year":"2010","journal-title":"Biol. Trace Elem. Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1007\/s11274-005-9041-0","article-title":"Purification and characterization of a sodium-stimulated \u03b2-galactosidasefrom Bifidobacterium longum CCRC 15708","volume":"22","author":"Hsu","year":"2006","journal-title":"World J. Microbiol. Biotechnol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"95","DOI":"10.2478\/BF02476241","article-title":"\u03b2-Galactosidase from Penicillium canescens. Properties and immobilization","volume":"3","author":"Budriene","year":"2005","journal-title":"Open Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"103442","DOI":"10.1016\/j.bioorg.2019.103442","article-title":"Purification and characterization of \u03b2-galactosidase from newly isolated Aspergillus terreus (KUBCF1306) and evaluating its efficacy on breast cancer cell line (MCF-7)","volume":"94","author":"Vidya","year":"2020","journal-title":"Bioorg. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"756","DOI":"10.1007\/s00216-003-1829-x","article-title":"Kinetic substrate quantification by fitting the enzyme reaction curve to the integrated Michaelis-Menten equation","volume":"375","author":"Liao","year":"2003","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Hildebrandt, P., Wanarska, M., and Kur, J. (2009). A new cold-adapted beta-D-galactosidase from the Antarctic Arthrobacter sp. 32c\u2014gene cloning, overexpression, purification and properties. BMC Microbiol., 9.","DOI":"10.1186\/1471-2180-9-151"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"19514","DOI":"10.1073\/pnas.1011100107","article-title":"Genome analysis of Bifidobacterium bifidum PRL2010 reveals metabolic pathways for host-derived glycan foraging","volume":"107","author":"Turroni","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4097","DOI":"10.1111\/febs.13908","article-title":"Crystal structure of \u03b2-1,6-galactosidase from Bifidobacterium bifidum S17: Trimeric architecture, molecular determinants of the enzymatic activity and its inhibition by \u03b1-galactose","volume":"283","author":"Godoy","year":"2016","journal-title":"FEBS J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1063","DOI":"10.1007\/s00253-020-11084-y","article-title":"Metagenomic identification, purification and characterisation of the Bifidobacterium adolescentis BgaC \u03b2-galactosidase","volume":"105","author":"Mulualem","year":"2021","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1093\/glycob\/cwt104","article-title":"Distinct substrate specificities of three glycoside hydrolase family 42 \u03b2-galactosidases from Bifidobacterium longum subsp. infantis ATCC 15697","volume":"24","author":"Viborg","year":"2014","journal-title":"Glycobiology"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.1111\/mmi.12815","article-title":"A \u03b21-6\/\u03b21-3 galactosidase from Bifidobacterium animalis subsp. lactis\u2005Bl-04 gives insight into sub-specificities of \u03b2-galactoside catabolism within Bifidobacterium","volume":"94","author":"Viborg","year":"2014","journal-title":"Mol. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1007\/s00284-009-9521-9","article-title":"Functional identification of a putative beta-galactosidase gene in the special lac gene cluster of Lactobacillus acidophilus","volume":"60","author":"Pan","year":"2010","journal-title":"Curr. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"8439","DOI":"10.1007\/s00253-019-10092-x","article-title":"A new \u03b2-galactosidase extracted from the infant feces with high hydrolytic and transgalactosylation activity","volume":"103","author":"Xin","year":"2019","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1007\/s00253-004-1745-9","article-title":"\u03b2-Galactosidase from Bifidobacterium adolescentis DSM20083 prefers \u03b2(1,4)-galactosides over lactose","volume":"66","author":"Hinz","year":"2004","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.enzmictec.2016.12.002","article-title":"Effect of protein load on stability of immobilized enzymes","volume":"98","author":"Pedrero","year":"2017","journal-title":"Enzym. Microb. Technol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/S0022-2836(02)00746-5","article-title":"Trimeric crystal structure of the glycoside hydrolase family 42 beta-galactosidase from Thermus thermophilus A4 and the structure of its complex with galactose","volume":"322","author":"Hidaka","year":"2002","journal-title":"J. Mol. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"649698","DOI":"10.3389\/fmicb.2021.649698","article-title":"Selective Isolation of Bifidobacterium From Human Faeces Using Pangenomics, Metagenomics, and Enzymology","volume":"12","author":"Yang","year":"2021","journal-title":"Front. Microbiol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"3126","DOI":"10.1021\/acs.biochem.7b01253","article-title":"Stereospecific Formation of Z-Trisubstituted Double Bonds by the Successive Action of Ketoreductase and Dehydratase Domains from trans-AT Polyketide Synthases","volume":"57","author":"Xie","year":"2018","journal-title":"Biochemistry"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1186\/s13068-017-0923-9","article-title":"A new method to evaluate temperature vs. pH activity profiles for biotechnological relevant enzymes","volume":"10","author":"Herlet","year":"2017","journal-title":"Biotechnol. Biofuels"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1385\/BTER:93:1-3:87","article-title":"Effects of metal ions on activity of plasmin","volume":"93","author":"Nowak","year":"2003","journal-title":"Biol. Trace Elem. Res."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1016\/j.ijbiomac.2020.04.045","article-title":"Pitfalls in the 3,5-dinitrosalicylic acid (DNS) assay for the reducing sugars: Interference of furfural and 5-hydroxymethylfurfural","volume":"156","author":"Deshavath","year":"2020","journal-title":"Int. J. Biol. Macromol."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/27\/14\/4497\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:50:08Z","timestamp":1760140208000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/27\/14\/4497"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,14]]},"references-count":44,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["molecules27144497"],"URL":"https:\/\/doi.org\/10.3390\/molecules27144497","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,14]]}}}