Team:Stuttgart/Laccasewiki-Experts
On this page we have collected all literature and experts that could be helpful or interesting for future laccase-based projects. We divided them by theme (English sources listed first, then German) and summarized the
core of the paper briefly for ease of use. If you have further questions about a German literature source, please do not hesitate to contact any of our teams, as we would be happy to provide you with the information you need for
your project to succeed!
Experts
- Dr. Sabine Sané
- Lecturer and researcher at the Institute for Microsystem Technology - IMTEK at Albert-Ludwigs-Universität Freiburg
- Expertise: Bio-electrochemical enzymatic systems to produce electricity and to degrade micropollutants in wastewater
- Find them here: https://www.biooekonomie-bw.de/fachbeitrag/aktuell/pilzsuppe-als-klaeranlage-und-biobrennstoffzelle
- Helped us with: Advice on laccase experimental conditions and laccase based system placement within a wastewater treatment plant, discussed her work with laccase that led to her winning the international Huber Technology Prize 2014: Future Water
- Dr. Dietmar Schlosser
- Group Leader Environmental Mycology at Helmholtz Center for Environmental Research - UFZ
- Expertise: Laccases
- Find them here: https://www.ufz.de/index.php?de=39080
- Helped us with: Advice on different algae laccases, best growing conditions and lab advice
- Prof. Dr. Antonio Pierik
- Professor of Biochemistry at Technical University of Kaiserslautern
- Expertise: Biochemistry and metallic proteins (such as laccase)
- Find them here: https://www.chemie.uni-kl.de/en/pierik/members-of-the-group/prof-dr-antonio-pierik/
- Helped us with: Proteins with metallic atoms such as laccase, gave recommendations for production in algae and how to avoid potential difficulties that could arise
- PhD Yunrong Chai
- Associate Professor of Biology at Northwestern University
- Expertise: Microbial genomics and biofilm formation
- Find them here: https://cos.northeastern.edu/people/yunrong-chai/
- Helped us with: Improvement of B. subtilis for our needs, fusion proteins with TasA
- Prof. Dr. Jörg Stülke
- Professor of Microbiology at the Institute for Microbiology and Genetics in Göttingen Graduate Center for Neurosciences, Biophysics, and Molecular Biosciences
- Expertise: Bacillus subtilis
- Find them here: https://www.uni-goettingen.de/en/58040.html
- Helped us with: Provided the tasA sinR knockout strain (GP1622), lab advice, advice on B. subtilis
- Prof. Dr. Susanne Lackner
- Professor in the department of wastewater management at the Technical University of Darmstadt
- Expertise: Microbial communities for wastewater purification and degradation of harmful pharmaceutical residues
- Find them here: https://www.iwar.tu-darmstadt.de/abwasserwirtschaft/fachgebiet_abwasserwirtschaft/mitarbeiter_8/Lackner.de.jsp
- Helped us with: Identifying harmful substances in wastewater, carrier material for implementation
- Prof. Dr. Jörg Oehlmann
- Professor of Aquatic ecotoxicology at Goethe University Frankfurt am Main
- Expertise: Micropollutants’ effects on aquatic environment
- Find them here: https://www.bio.uni-frankfurt.de/45735703/Aquatische_%C3%96kotoxikologie
- Helped us with: Identifying harmful substances in water, implementation in wastewater treatment plants (carrier material)
- Dr. Patrick Schröder
- Research assistant at Federal Environment Agency
- Expertise: Biodegradation of pharmaceuticals and development of antimicrobial resistances
- Find them here: https://www.umweltbundesamt.de/impressum
- Helped us with: Provided understanding of dangers caused by pharmaceuticals and prevention of unnecessary water toxification through science communication
- Dr. Bertram Kuch
- Institute Sanitary Engineering, Water Quality and Solid Waste management University of Stuttgart
- Expertise: Head of the department for the analysis of micropollutants
- Find them here: https://www.iswa.uni-stuttgart.de/institut/team/Kuch-00001/
- Helped us with: Decision on what substrates we should focus on degrading with our laccases as well as theoretical background for the establishment of an analytical method
- Dr.-Ing. Henning Knerr
- Manager of tectra and Division manager of wastewater treatment and water quality at Technical University of Kaiserslautern
- Expertise: Engineering and wastewater management
- Find them here: https://www.bauing.uni-kl.de/en/wir/team/leadership/dr-ing-henning-knerr/
- Helped us with: Understanding major micropollutants in Germany (Diclofenac specific) and Rheinland-Pfalz wastewater treatment plant design and smaller treatment plant methods of cleaning micropollutants
- Dr. Gerd Maack
- Environmental Risk Assessment of Pharmaceuticals in German Environment Agency (UBA), EDA-EMERGE Advisory Board Member
- Expertise: German micropollutants and environmental effects
- Find them here: https://www.umweltbundesamt.de/en/topics/chemicals/pharmaceuticals https://www.ufz.de/eda-emerge/index.php?en=30284
- Helped us with: Understanding major micropollutants in Germany (Diclofenac specific) and worldwide environmental effects
- Dipl. Ing. Udo Bäuerle
- Construction Engineer, co-owner of “B&P Beratende Ingenieure”
- Expertise: Water management and wastewater treatment
- Find them here: http://www.bp-bi.de/ger/pages/personal01.htm
- Helped us with: Explanation of wastewater purification, implementation of a biofilm in wastewater treatment plants
- Prof. Dr. Karl-Heinrich Engesser
- Institute of Sanitary Engineering, Water Quality and Solid Waste Management University of Stuttgart
- Expertise: Wastewater management and biocatalysis
- Find them here: https://www.iswa.uni-stuttgart.de/en/institute/team/Engesser-00004/
- Helped us with: The implementation possibilities of our project in a wastewater plant and possible difficulties and what would be required to overcome them
- Christian Kaiser
- Member of the Initiative Progressive Agrarwende
- Expertise: Biology student at the University of Bonn, focus in pharmaceutical biology on bacterial natural substances and their biosynthesis
- Find them here: https://progressive-agrarwende.org/ueber-uns/
- Helped us with: Our project’s design and realistic implementation, discussion on GMO regulations and micropollutants generally
- Mike Short
- Village of Archbold Wastewater Superintendent
- Expertise: American Wastewater Treatment plant
- Find them here: http://archbold.com/wastewater/
- Helped us with: Comparisons between US wastewater treatment and German wastewater treatment methods
- Dan Avers
- Village of Archbold Water Distribution Chief
- Expertise: American Water treatment plant
- Find them here:
- Helped us with: Comparisons between US water treatment and German water treatment methods
- Florian Heyn
- Management of wastewater treatment plant in Alzenau, Germany
- Expertise: German wastewater treatment plant
- Find them here: https://www.alzenau.de/B%C3%BCrger/Rathaus-Service/Verwaltung/Mitarbeiter-von-A-Z/index.php?ModID=9&object=tx%7C2413.11&FID=2413.148.1&NavID=2413.39&La=1
- Helped us with: Understanding how a German wastewater treatment plant operates
- Thomas Seeger
- Management of wastewater treatment plant in Weiterstadt, Germany
- Expertise: German wastewater treatment plant
- Find them here: https://www.weiterstadt.de/verwaltung-service/rathaus/telefonverzeichnis/index.php?we_objectID=1403
- Helped us with: Understanding how a German wastewater treatment plant operates
- Dr. Ulrich Ehlers
- Head of Unit 403 “Deliberate Release and Placing on the Market” of 'Genetic Engineering' department of the Federal Office of Consumer Protection and Food Safety (BVL)
- Expertise: Genetic engineering regulations in Germany
- Find them here: https://www.bvl.bund.de/EN/Tasks/06_Genetic_engineering/genetic_engineering_node.html
- Helped us with: Understanding wastewater treatment plant restrictions for GMO in Germany and bioreactor specifications
- Prof. Dr. Sibylle Gaisser
- Professor of Industrial Biotechnology at Hochschule Ansbach
- Expertise: Socio-economic analysis and technology assessment
- Find them here: https://www.hs-ansbach.de/personen/gaisser-sibylle/
- Helped us with: Assessment of risks, advice on complying with ethical standards and safety concerns.
- Expertise: Socio-economic analysis and technology assessment
- Prof. Dr. Alfred Nordmann
- Professor of philosophy and history of the sciences and technosciences at the Technical University of Darmstadt
- Expertise: Ethics
- Find them here: https://www.philosophie.tu-darmstadt.de/institut_phil/mitarbeiter_innen_phil/professoren/a_nordmann/biography_2.de.jsp
- Helped us with: Introduction into responsible research and innovation, importance of science communication
- Expertise: Ethics
Arregui, L.; Ayala, M.; Gómez-Gil, X.; Gutiérrez-Soto, G. Laccases: Structure, Function, and Potential Application in Water Bioremediation. Microbial
Cell Factories. BioMed Central Ltd. November 14, 2019, pp 1–33.DOI:10.1186/s12934-019-1248-0.
This review compares many fungal and bacterial laccases and their potential for use in wastewater treatment.
Mate, D. M.; Alcalde, M. Research Review Paper Laccase Engineering: From Rational Design to Directed Evolution. Biotechnol. Adv. 2014, 33, 25–40. DOI:10.1016/j.biotechadv.2014.12.007.
Many different laccases, both fungal and bacterial, are compared and it discusses how these could be modified to improve effectiveness in reactions.
Margot, J.; Bennati-Granier, C.; Maillard, J. Bacterial versus Fungal Laccase: Potential for Micropollutant Degradation; 2013; Vol. 3. DOI:/10.1186/2191-0855-3-63.
Evaluation and comparison of laccase-producing organisms, particularly useful for the comparison of T. versicolor and S. cyaneus.
Kittl R, Mueangtoom K, Gonaus C, A chloride tolerant laccase from the plant pathogen ascomycete Botrytis aclada expressed at high levels in Pichia pastoris. J Biotechnol. 2012 Jan 20;157(2):304-14. doi: 10.1016/j.jbiotec.2011.11.021. Epub 2011 Dec 9. PMID: 22178779. DOI:10.1016/j.jbiotec.2011.11.02.
Botrytis aclada laccase (BaLac) is discussed at length in this paper as a viable option for a laccase with a low pI and chloride tolerant construct, factors which normally limit laccase effectiveness.
Scheiblbrandner, S., Breslmayr, E., Csarman, F. Evolving stability and pH-dependent activity of the high redox potential Botrytis aclada laccase for enzymatic fuel cells. Sci Rep 7, 13688 (2017). DOI:10.1038/s41598-017-13734-0.
Experiments that delve into increasing the pH optimum of Botrytis aclada laccase (BaLac), a laccase with a low pI and chloride tolerant construct, factors which normally limit laccase effectiveness.
Yang Q, Zhang M, Zhang M Characterization of a Novel, Cold-Adapted, and Thermostable Laccase-Like Enzyme With High Tolerance for Organic Solvents and Salt and Potent Dye Decolorization Ability, Derived From a Marine Metagenomic Library. Front Microbiol. 2018;9:2998. Published 2018 Dec 5. DOI:10.3389/fmicb.2018.02998
Investigation over the marine laccase know as Lac1326, an extremely thermostable laccase construct.
Zeng, J.; Zhu, Q.; Wu, Y. Oxidation of Polycyclic Aromatic Hydrocarbons Using Bacillus Subtilis CotA with High Laccase Activity and Copper Independence. Chemosphere 2016, 148, 1–7. DOI:10.1016/j.chemosphere.2016.01.019.
The bacterial laccases CueO and CotA are compared regarding the oxidation rates of 15 different polycyclic aromatic hydrocarbons.
German literature:
Ricklefs, E. Charakterisierung Einer Neuen Bakteriellen Laccase Und Deren Anwendung in Einer Multi-Enzymatischen Kaskade Zur Synthese von Lignanen. https://docserv.uni-duesseldorf.de/servlets/DerivateServlet/Derivate-41000/Ricklefs%2C%20Esther_Doktorarbeit_pdf_a1.pdf (accessed on Oct 13, 2020)
A new bacterial laccase from Corynebacterium glutamicum has been characterized and analysed. Furthermore bacterial laccases were used for multi-enzymatic cascades for fine chemical production.
This review compares many fungal and bacterial laccases and their potential for use in wastewater treatment.
Mate, D. M.; Alcalde, M. Research Review Paper Laccase Engineering: From Rational Design to Directed Evolution. Biotechnol. Adv. 2014, 33, 25–40. DOI:10.1016/j.biotechadv.2014.12.007.
Many different laccases, both fungal and bacterial, are compared and it discusses how these could be modified to improve effectiveness in reactions.
Margot, J.; Bennati-Granier, C.; Maillard, J. Bacterial versus Fungal Laccase: Potential for Micropollutant Degradation; 2013; Vol. 3. DOI:/10.1186/2191-0855-3-63.
Evaluation and comparison of laccase-producing organisms, particularly useful for the comparison of T. versicolor and S. cyaneus.
Kittl R, Mueangtoom K, Gonaus C, A chloride tolerant laccase from the plant pathogen ascomycete Botrytis aclada expressed at high levels in Pichia pastoris. J Biotechnol. 2012 Jan 20;157(2):304-14. doi: 10.1016/j.jbiotec.2011.11.021. Epub 2011 Dec 9. PMID: 22178779. DOI:10.1016/j.jbiotec.2011.11.02.
Botrytis aclada laccase (BaLac) is discussed at length in this paper as a viable option for a laccase with a low pI and chloride tolerant construct, factors which normally limit laccase effectiveness.
Scheiblbrandner, S., Breslmayr, E., Csarman, F. Evolving stability and pH-dependent activity of the high redox potential Botrytis aclada laccase for enzymatic fuel cells. Sci Rep 7, 13688 (2017). DOI:10.1038/s41598-017-13734-0.
Experiments that delve into increasing the pH optimum of Botrytis aclada laccase (BaLac), a laccase with a low pI and chloride tolerant construct, factors which normally limit laccase effectiveness.
Yang Q, Zhang M, Zhang M Characterization of a Novel, Cold-Adapted, and Thermostable Laccase-Like Enzyme With High Tolerance for Organic Solvents and Salt and Potent Dye Decolorization Ability, Derived From a Marine Metagenomic Library. Front Microbiol. 2018;9:2998. Published 2018 Dec 5. DOI:10.3389/fmicb.2018.02998
Investigation over the marine laccase know as Lac1326, an extremely thermostable laccase construct.
Zeng, J.; Zhu, Q.; Wu, Y. Oxidation of Polycyclic Aromatic Hydrocarbons Using Bacillus Subtilis CotA with High Laccase Activity and Copper Independence. Chemosphere 2016, 148, 1–7. DOI:10.1016/j.chemosphere.2016.01.019.
The bacterial laccases CueO and CotA are compared regarding the oxidation rates of 15 different polycyclic aromatic hydrocarbons.
German literature:
Ricklefs, E. Charakterisierung Einer Neuen Bakteriellen Laccase Und Deren Anwendung in Einer Multi-Enzymatischen Kaskade Zur Synthese von Lignanen. https://docserv.uni-duesseldorf.de/servlets/DerivateServlet/Derivate-41000/Ricklefs%2C%20Esther_Doktorarbeit_pdf_a1.pdf (accessed on Oct 13, 2020)
A new bacterial laccase from Corynebacterium glutamicum has been characterized and analysed. Furthermore bacterial laccases were used for multi-enzymatic cascades for fine chemical production.
Alharbi, S. K.; Nghiem, L. D.; van de Merwe, J. P. Degradation of Diclofenac, Trimethoprim, Carbamazepine, and Sulfamethoxazole by Laccase from
Trametes Versicolor: Transformation Products and Toxicity of Treated Effluent. Biocatal. Biotransformation 2019, 37 (6), 399–408. DOI:10.1080/10242422.2019.1580268 .
Degradation products of diclofenac and carbamazepine with the laccase of Trametes versicolor discussed, as well as HPLC method.
Jones, S. M.; Solomon, E. I. Electron Transfer and Reaction Mechanism of Laccases. Cellular and Molecular Life Sciences. Birkhauser Verlag AG March 1, 2015, pp 869–883. DOI:10.1007/s00018-014-1826-6.
The electron transfer process and the O2 reduction in laccases is outlined.
Naghdi, M.; Taheran, M.; Brar, S. K. Removal of Pharmaceutical Compounds in Water and Wastewater Using Fungal Oxidoreductase Enzymes. Environmental Pollution. Elsevier Ltd March 1, 2018, pp 190–213. DOI:10.1016/j.envpol.2017.11.060.
This review summarizes and analyzes the studies performed on pharmaceutically active compounds in wastewater using oxidoreductase enzymes for their degradation. The review also offers an insight into enzymes immobilization, fungal reactors, mediators, degradation mechanisms and transformation products.
Erythromycin Esterase Type II (EreB):
Morar, M.; Pengelly, K.; Koteva, K. Mechanism and Diversity of the Erythromycin Esterase Family of Enzymes. 2012, 51, 1740–1751. DOI:10.1021/bi201790u.
The mechanisms of the Erythromycin and Azithromycin inactivation by the enzyme EreB are described.This takes place by the enzymatic hydrolysis of the macrolactone ring.
Degradation products of diclofenac and carbamazepine with the laccase of Trametes versicolor discussed, as well as HPLC method.
Jones, S. M.; Solomon, E. I. Electron Transfer and Reaction Mechanism of Laccases. Cellular and Molecular Life Sciences. Birkhauser Verlag AG March 1, 2015, pp 869–883. DOI:10.1007/s00018-014-1826-6.
The electron transfer process and the O2 reduction in laccases is outlined.
Naghdi, M.; Taheran, M.; Brar, S. K. Removal of Pharmaceutical Compounds in Water and Wastewater Using Fungal Oxidoreductase Enzymes. Environmental Pollution. Elsevier Ltd March 1, 2018, pp 190–213. DOI:10.1016/j.envpol.2017.11.060.
This review summarizes and analyzes the studies performed on pharmaceutically active compounds in wastewater using oxidoreductase enzymes for their degradation. The review also offers an insight into enzymes immobilization, fungal reactors, mediators, degradation mechanisms and transformation products.
Erythromycin Esterase Type II (EreB):
Morar, M.; Pengelly, K.; Koteva, K. Mechanism and Diversity of the Erythromycin Esterase Family of Enzymes. 2012, 51, 1740–1751. DOI:10.1021/bi201790u.
The mechanisms of the Erythromycin and Azithromycin inactivation by the enzyme EreB are described.This takes place by the enzymatic hydrolysis of the macrolactone ring.
Patel, S.; Kalia, V. C.; Choi, J.-H. Immobilization of Laccase on SiO2 Nanocarriers Improves Its Stability and Reusability Biological Hydrogen
Production, Polyhydroxyalkanoates, Quorum Sensing Inhibitors, Bacterial Identification View Project INJM Special ISSUE View Project. Artic. J. Microbiol. Biotechnol. 2014. DOI:10.4014/jmb.1401.01025.
Immobilization of laccase on SiO2 nanoparticles to overcome problems associated with stability and reusability of the free enzyme is presented.
Huang, J.; Liu, S.; Zhang, C. Programmable and Printable Bacillus Subtilis Biofilms as Engineered Living Materials. Nat. Chem. Biol. 2019. DOI:10.1038/s41589-018-0169-2.
Immobilization of proteins by producing a fusion protein with the major protein component (TasA) of Bacillus subtilis biofilm is shown.
German literature:
Dreifke, M.; Fröba, M. Immobilisierung von Enzymen: Spielerei Oder Biotechnologischer Fortschritt? Springer 2017, 23 (1), 95–97. DOI:10.1007/s12268-017-0769-5.
Different enzyme immobilization strategies especially about the enzyme immobilization on silica foam are explained and compared.
Immobilization of laccase on SiO2 nanoparticles to overcome problems associated with stability and reusability of the free enzyme is presented.
Huang, J.; Liu, S.; Zhang, C. Programmable and Printable Bacillus Subtilis Biofilms as Engineered Living Materials. Nat. Chem. Biol. 2019. DOI:10.1038/s41589-018-0169-2.
Immobilization of proteins by producing a fusion protein with the major protein component (TasA) of Bacillus subtilis biofilm is shown.
German literature:
Dreifke, M.; Fröba, M. Immobilisierung von Enzymen: Spielerei Oder Biotechnologischer Fortschritt? Springer 2017, 23 (1), 95–97. DOI:10.1007/s12268-017-0769-5.
Different enzyme immobilization strategies especially about the enzyme immobilization on silica foam are explained and compared.
Following the Flow An Inside Look
at Wastewater Treatment https://www.wef.org/globalassets/assets-wef/3---resources/for-the-public/public-information/following-the-flow-book-an-inside-look-at-wastewater-treatment.pdf (accessed Oct 1, 2020).
There is general information about Wastewater Treatment Plant structures as an initial overview described by the Water Environment Federation (WEF).
Pistocchi, A.; Dorati, C.; Grizzetti, B.; Udias, A.; Vigiak, O.; Zanni, M. Water Quality in Europe : Effects of the Urban Wastewater Treatment Directive; 2019. DOI:10.2760/303163.
This report shows what micropollutants can be found in wastewater and whether they are transformed in wastewater treatment.
German literature:
Arzneimittelwirkstoffe | Umweltbundesamt https://www.umweltbundesamt.de/themen/wasser/fluesse/zustand/arzneimittelwirkstoffe#undefined (accessed Sep 3, 2020).
Problematic pollutants in German wastewater are described in this article.
There is general information about Wastewater Treatment Plant structures as an initial overview described by the Water Environment Federation (WEF).
Pistocchi, A.; Dorati, C.; Grizzetti, B.; Udias, A.; Vigiak, O.; Zanni, M. Water Quality in Europe : Effects of the Urban Wastewater Treatment Directive; 2019. DOI:10.2760/303163.
This report shows what micropollutants can be found in wastewater and whether they are transformed in wastewater treatment.
German literature:
Arzneimittelwirkstoffe | Umweltbundesamt https://www.umweltbundesamt.de/themen/wasser/fluesse/zustand/arzneimittelwirkstoffe#undefined (accessed Sep 3, 2020).
Problematic pollutants in German wastewater are described in this article.
Teams that used laccase for inactivation of micro pollutants:
Team TU Munich in 2013:
They used a laccase from Bacillus pumilus and EreB from Escherichia coli for biodegradation of Xenobiotics.
Team Bielefeld-Germany in 2012:
They used five different laccases and even compared them to each other to clean wastewater from synthetic estrogens and aromatic compounds.
Team Stockholm in 2018:
They used the laccase from Trametes versicolor, immobilized on magnetic beads, to inactivate sulfamethoxazole.
Team Western Canada in 2019:
They also used the laccase from Trametes versicolor and immobilized it via the spy-tag/spy-catcher-system on the major curlin subunit (CsgA) of E. coli to clean wastewater from emerging compounds.
Teams that used laccase for other targets:
Team Saint Joseph in 2019:
They used the laccase from Trametes versicolor for dye-decolorization.
Team Edinburgh OG in 2019:
They used the laccase from Bacillus pumilus for dye-decolorization.
Team Hong Kong HKUST in 2018:
They used the laccase from Escherichia coli for poly-ethylen degradation.
Team British Columbia in 2016:
They displayed small laccases on the cell surface of C. crescentus for degradation of lignin cellulose.
Team SHSBNU China in 2018:
They used the CotA laccase from Bacillus subtilis and immobilized it via spy-tag/spy-catcher-system on the major curlin subunit (CsgA) of E. coli for dye-decolorization.
Team Paris Bettencourt in 2016:
They used six enzymes, among which the laccase from Bacillus pumilus to remove stains made by Anthocyanins.
Team TU Munich in 2013:
They used a laccase from Bacillus pumilus and EreB from Escherichia coli for biodegradation of Xenobiotics.
Team Bielefeld-Germany in 2012:
They used five different laccases and even compared them to each other to clean wastewater from synthetic estrogens and aromatic compounds.
Team Stockholm in 2018:
They used the laccase from Trametes versicolor, immobilized on magnetic beads, to inactivate sulfamethoxazole.
Team Western Canada in 2019:
They also used the laccase from Trametes versicolor and immobilized it via the spy-tag/spy-catcher-system on the major curlin subunit (CsgA) of E. coli to clean wastewater from emerging compounds.
Teams that used laccase for other targets:
Team Saint Joseph in 2019:
They used the laccase from Trametes versicolor for dye-decolorization.
Team Edinburgh OG in 2019:
They used the laccase from Bacillus pumilus for dye-decolorization.
Team Hong Kong HKUST in 2018:
They used the laccase from Escherichia coli for poly-ethylen degradation.
Team British Columbia in 2016:
They displayed small laccases on the cell surface of C. crescentus for degradation of lignin cellulose.
Team SHSBNU China in 2018:
They used the CotA laccase from Bacillus subtilis and immobilized it via spy-tag/spy-catcher-system on the major curlin subunit (CsgA) of E. coli for dye-decolorization.
Team Paris Bettencourt in 2016:
They used six enzymes, among which the laccase from Bacillus pumilus to remove stains made by Anthocyanins.