{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,10]],"date-time":"2026-03-10T10:01:11Z","timestamp":1773136871692,"version":"3.50.1"},"reference-count":38,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2021,12,14]],"date-time":"2021-12-14T00:00:00Z","timestamp":1639440000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100009708","name":"Novo Nordisk Foundation","doi-asserted-by":"publisher","award":["NNF20SA0066233"],"award-info":[{"award-number":["NNF20SA0066233"]}],"id":[{"id":"10.13039\/501100009708","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Energies"],"abstract":"<jats:p>This study evaluated the individual and combined effects of inhibitory compounds formed during pretreatment of lignocellulosic biomass on the growth of Bacillus subtilis. Ten inhibitory compounds commonly present in lignocellulosic hydrolysates were evaluated, which included sugar degradation products (furfural and 5-hydroxymethylfurfural), acetic acid, and seven phenolic compounds derived from lignin (benzoic acid, vanillin, vanillic acid, ferulic acid, p-coumaric acid, 4-hydroxybenzoic acid, and syringaldehyde). For the individual inhibitors, syringaldehyde showed the most toxic effect, completely inhibiting the strain growth at 0.1 g\/L. In the sequence, assays using mixtures of the inhibitory compounds at a concentration of 12.5% of their IC50 value were performed to evaluate the combined effect of the inhibitors on the strain growth. These experiments were planned according to a Plackett\u2013Burman experimental design. Statistical analysis of the results revealed that in a mixture, benzoic acid and furfural were the most potent inhibitors affecting the growth of B. subtilis. These results contribute to a better understanding of the individual and combined effects of inhibitory compounds present in biomass hydrolysates on the microbial performance of B. subtilis. Such knowledge is important to advance the development of sustainable biomanufacturing processes using this strain cultivated in complex media produced from lignocellulosic biomass, supporting the development of efficient bio-based processes using B. subtilis.<\/jats:p>","DOI":"10.3390\/en14248419","type":"journal-article","created":{"date-parts":[[2021,12,14]],"date-time":"2021-12-14T09:34:25Z","timestamp":1639474465000},"page":"8419","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["Effects of Inhibitory Compounds Present in Lignocellulosic Biomass Hydrolysates on the Growth of Bacillus subtilis"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2468-7850","authenticated-orcid":false,"given":"Lucas","family":"van der Maas","sequence":"first","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, S\u00f8ltofts Plads, Building 223, 2800 Kongens Lyngby, Denmark"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1445-4770","authenticated-orcid":false,"given":"Jasper L. S. P.","family":"Driessen","sequence":"additional","affiliation":[{"name":"Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, Building 220, 2800 Kongens Lyngby, Denmark"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7182-6198","authenticated-orcid":false,"given":"Solange I.","family":"Mussatto","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, S\u00f8ltofts Plads, Building 223, 2800 Kongens Lyngby, Denmark"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1111\/gcbb.12586","article-title":"Integrated lignocellulosic value chains in a growing bioeconomy: Status quo and perspectives","volume":"11","author":"Dahmen","year":"2018","journal-title":"GCB Bioenergy"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"111620","DOI":"10.1016\/j.rser.2021.111620","article-title":"New trends in bioprocesses for lignocellulosic biomass and CO2 utilization","volume":"152","author":"Mussatto","year":"2021","journal-title":"Renew. Sustain. Energy Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"122847","DOI":"10.1016\/j.biortech.2020.122847","article-title":"Innovation and strategic orientations for the development of advanced biorefineries","volume":"302","author":"Dragone","year":"2020","journal-title":"Bioresour. Technol."},{"key":"ref_4","unstructured":"BCC Research (2021, October 23). Global Markets for Enzymes in Industrial Applications. Available online: https:\/\/www.bccresearch.com\/market-research\/biotechnology\/global-markets-for-enzymes-in-industrial-applications.html."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Ravindran, R., Hassan, S.S., Williams, G.A., and Jaiswal, A.K. (2018). A review on bioconversion of agro-industrial wastes to industrially important enzymes. Bioengineering, 5.","DOI":"10.3390\/bioengineering5040093"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Mussatto, S.I. (2016). Biomass pretreatment, biorefineries, and potential products for a bioeconomy development. Biomass Fractionation Technologies for a Lignocellulosic Feedstock Based Biorefinery, Elsevier.","DOI":"10.1016\/B978-0-12-802323-5.00001-3"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.rser.2017.03.091","article-title":"Fermentative hydrogen production using lignocellulose biomass: An overview of pre-treatment methods, inhibitor effects and detoxification experiences","volume":"77","author":"Sivagurunathan","year":"2017","journal-title":"Renew. Sustain. Energy Rev."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.biortech.2003.10.005","article-title":"Alternatives for detoxification of diluted-acid lignocellulosic hydrolyzates for use in fermentative processes: A review","volume":"93","author":"Mussatto","year":"2004","journal-title":"Bioresour. Technol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"113799","DOI":"10.1016\/j.indcrop.2021.113799","article-title":"Effects of inhibitory compounds derived from lignocellulosic biomass on the growth of the wild-type and evolved oleaginous yeast Rhodosporidium toruloides","volume":"170","author":"Liu","year":"2021","journal-title":"Ind. Crops Prod."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1139\/w03-076","article-title":"Developments in the use of Bacillus species for industrial production","volume":"50","author":"Schallmey","year":"2004","journal-title":"Can. J. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Hemmerich, J., Wiechert, W., and Oldiges, M. (2017). Automated growth rate determination in high-throughput microbioreactor systems. BMC Res. Notes, 10.","DOI":"10.1186\/s13104-017-2945-6"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1016\/0740-0020(90)90029-H","article-title":"A mathematical approach toward defining and calculating the duration of the lag phase","volume":"7","author":"Buchanan","year":"1990","journal-title":"Food Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2189","DOI":"10.1128\/jb.179.7.2189-2193.1997","article-title":"Mutations affecting substrate specificity of the Bacillus subtilis multidrug transporter Bmr","volume":"179","author":"Klyachko","year":"1997","journal-title":"J. Bacteriol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/S0304-3800(01)00271-X","article-title":"Plackett-Burman technique for sensitivity analysis of many-parametered models","volume":"141","author":"Beres","year":"2001","journal-title":"Ecol. Model."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1016\/0165-7992(89)90125-5","article-title":"Specificity of the interaction of furfural with DNA","volume":"225","author":"Hadi","year":"1989","journal-title":"Mutat. Res. Lett."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"769","DOI":"10.1042\/bj3630769","article-title":"Inhibition effects of furfural on alcohol dehydrogenase, aldehyde dehydrogenase and pyruvate dehydrogenase","volume":"363","author":"Modig","year":"2002","journal-title":"Biochem. J."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1002\/(SICI)1097-0290(19991005)65:1<24::AID-BIT4>3.0.CO;2-2","article-title":"Effect of selected aldehydes on the growth and fermentation of ethanologenic Escherichia coli","volume":"65","author":"Zaldivar","year":"1999","journal-title":"Biotechnol. Bioeng."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/0032-9592(91)80003-8","article-title":"Utilization of sugar cane bagasse hemicellulosic hydrolysate by Pichia stipitis for the production of ethanol","volume":"26","author":"Roberto","year":"1991","journal-title":"Process Biochem."},{"key":"ref_19","first-page":"727","article-title":"Biodegradation of furfural by Bacillus subtilis strain DS3","volume":"36","author":"Zheng","year":"2015","journal-title":"J. Environ. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1016\/j.biortech.2014.02.128","article-title":"An efficient process for lactic acid production from wheat straw by a newly isolated Bacillus coagulans strain IPE22","volume":"158","author":"Zhang","year":"2014","journal-title":"Bioresour. Technol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"9069","DOI":"10.1007\/s00253-016-7642-1","article-title":"Growth inhibition of S. cerevisiae, B. subtilis, and E. coli by lignocellulosic and fermentation products","volume":"100","author":"Pereira","year":"2016","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3604","DOI":"10.1128\/aem.61.10.3604-3608.1995","article-title":"The relationship between viability and intracellular pH in the yeast Saccharomyces cerevisiae","volume":"61","author":"Imai","year":"1995","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1007\/BF00170063","article-title":"Activity of glycolytic enzymes of Saccharomyces cerevisiae in the presence of acetic acid","volume":"34","author":"Pampulha","year":"1990","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1099\/00221287-78-2-261","article-title":"Control of metabolite secretion in Bacillus subtilis","volume":"78","author":"Speck","year":"1973","journal-title":"J. Gen. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/0141-0229(91)90193-E","article-title":"Acetic acid inhibition of d-xylose fermentation by Pichia stipitis","volume":"13","author":"Prior","year":"1991","journal-title":"Enzyme Microb. Technol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1002\/jobm.3620350309","article-title":"Effect of acetic acid on xylose fermentation to xylitol by Candida guilliermondii","volume":"35","author":"Felipe","year":"1995","journal-title":"J. Basic Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1002\/(SICI)1097-0290(19990405)63:1<46::AID-BIT5>3.0.CO;2-J","article-title":"Main and interaction effects of acetic acid, furfural, and p- hydroxybenzoic acid on growth and ethanol productivity of yeasts","volume":"63","author":"Palmqvist","year":"1999","journal-title":"Biotechnol. Bioeng."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2091","DOI":"10.1128\/aem.54.8.2091-2095.1988","article-title":"Effect of Benzoic Acid on Growth Yield of Yeasts Differing in Their Resistance to Preservatives","volume":"54","author":"Warth","year":"1988","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1201\/9781420028737.ch2","article-title":"Sodium benzoate and benzoic acid","volume":"Volume 145","author":"Chipley","year":"2005","journal-title":"Food Science and Technology"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"598","DOI":"10.7150\/ijbs.6091","article-title":"Molecular adaptation mechanisms employed by ethanologenic bacteria in response to lignocellulose-derived inhibitory compounds","volume":"9","author":"Ibraheem","year":"2013","journal-title":"Int. J. Biol. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"9579","DOI":"10.1007\/s00253-014-6158-9","article-title":"By-products resulting from lignocellulose pretreatment and their inhibitory effect on fermentations for (bio)chemicals and fuels","volume":"98","author":"Bakker","year":"2014","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1186\/s13068-019-1610-9","article-title":"Physiological mechanism of improved tolerance of Saccharomyces cerevisiae to lignin-derived phenolic acids in lignocellulosic ethanol fermentation by short-term adaptation","volume":"12","author":"Gu","year":"2019","journal-title":"Biotechnol. Biofuels"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1035","DOI":"10.1007\/s00253-009-1925-8","article-title":"Detoxification of model phenolic compounds in lignocellulosic hydrolysates with peroxidase for butanol production from Clostridium beijerinckii","volume":"83","author":"Cho","year":"2009","journal-title":"Appl Microbiol. Biotechnol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"275","DOI":"10.17221\/132\/2010-CJFS","article-title":"Antimicrobial and antioxidant properties of phenolic acids alkyl esters","volume":"28","author":"Merkl","year":"2010","journal-title":"Czech J. Food Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1378","DOI":"10.1111\/j.1365-2621.1983.tb09243.x","article-title":"Antibacterial activity of selected hydroxycinnamic acids","volume":"48","author":"Herald","year":"1983","journal-title":"J. Food Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2273","DOI":"10.1271\/bbb.62.2273","article-title":"Antimicrobial activity of 4-hydroxybenzoic acid and trans 4-hydroxycinnamic acid isolated and identified from rice hull","volume":"62","author":"Cho","year":"1998","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"9151","DOI":"10.1007\/s00253-014-6106-8","article-title":"Microbial inhibitors: Formation and effects on acetone-butanol-ethanol fermentation of lignocellulosic biomass","volume":"98","author":"Baral","year":"2014","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1186\/1754-6834-6-99","article-title":"Inhibition of growth of Zymomonas mobilis by model compounds found in lignocellulosic hydrolysates","volume":"6","author":"Franden","year":"2013","journal-title":"Biotechnol. Biofuels"}],"container-title":["Energies"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1996-1073\/14\/24\/8419\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:47:12Z","timestamp":1760168832000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1996-1073\/14\/24\/8419"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,14]]},"references-count":38,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["en14248419"],"URL":"https:\/\/doi.org\/10.3390\/en14248419","relation":{},"ISSN":["1996-1073"],"issn-type":[{"value":"1996-1073","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,12,14]]}}}