Research Group n. - CIRCC
Transcript
Research Group n. - CIRCC
Research Group n. 2 UNIVERSITA’ DEGLI STUDI DI BARI Cheap and environmentally sustainable stereoselective arylketones reduction by Lactobacillus reuteri whole cells Gennaro Agrimi1, Palmieri Luigi1, Isabella Pisano1, Paola Vitale2, Filippo Perna2, Antonio Scilimati2 1 Department of Biosciences, Biotechnology and Biopharmaceutics, University of Bari, via Orabona 4, 70124 Bari 2 Department of Pharmacy & Pharmaceutical Sciences, University of Bari "A. Moro", Via Orabona 4, 70125 Bari, Italy [email protected] Various Lactobacillus reuteri strains were investigated as whole cell catalysts for the bioreduction of acetophenone into optically active (R)-1-phenylethanol. L. reuteri DSM 20016 strain gave an almost complete substrate conversion, in a short reaction time and enantiomeric excess up to 99%. The acetophenone bioreduction was used as a model reaction to optimize temperature and reducing equivalent source (glucose, lactose, cheese way and lignocellulosic hydrolysates) to accomplish the biotransformation. The reduction of acetophenones into optically active (R)-1-arylethanols was also exploited to study L. reuteri DSM 20016 substrate specificity. In most of the cases, optically active (R)1-arylethanols have been obtained with both excellent chemical and optical yields and with (R)enantiopreference, through a cheap, simple and efficient process. Key words: Lactobacillus reuteri, bioreduction, whole cells catalysis Cheese whey: a promising raw material in biotechnological industrial fermentation processesImproved Saccharomyces Cerevisiae Growth on Cheese Whey by Controlling Enzymatic Lactose Hydrolysis Isabella Pisano, Gennaro Agrimi, Palmieri Luigi Department of Biosciences, Biotechnology and Biopharmaceutics, University of Bari, via Orabona 4, 70124 Bari [email protected] Disposal of whey generated in cheese industry is an environmental problem but could represent also an opportunity for the diary sector. A “bio-refinery-oriented“ approach for the valorization of cheese whey is a complex process involving multiple technologies that, if employed in an integrated way, might lead to the obtainment of value added products (biomass fine or bulk chemicals). This work is focused on the study and optimization of a fermentation process using whey as growth medium and carbon source. Lactose which is abundant in whey is an interesting carbon source. However, the microorganism more widely used in industrial fermentation processes, the yeast Saccharomyces cerevisiae, does not have a lactose metabolization system. We set up an innovative biotechnological process for the production on large scale of a not-genetically modified yeast biomass that can be used in different contexts, such as bread making, production of probiotics, nutraceuticals, bio-active molecules. In order to use the cheese whey as raw material for the cultivation of S. cerevisiae and to overcome the limitations in the use of lactose we used and externally added β-galactosidase. The careful optimization of the amount of added enzyme allowed the gradual release by hydrolysis and the simultaneous consumption of glucose and galactose with a consequent decrease of ethanol and increase of the biomass produced. Key words: whey, yeast fermentation, animal feed, biomass. Pubblications: 1) Perna F.M., Ricci M.A., Scilimati A., Mena M.C., Pisano I., Palmieri L., Agrimi G., Vitale P. (2016). Cheap and environmentally sustainable stereoselective arylketones reduction by Lactobacillus reuteri whole cells. JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, vol. 124, p. 29-37, ISSN: 1381-1177, doi: 10.1016/j.molcatb.2015.11.025 2) Pisano I., Agrimi G., Grosso G., Mena M.C., Ricci M.A., Palmieri L. (2015). Improved Saccharomyces Cerevisiae growth on cheese whey by controlling enzymatic lactose hydrolysis. CHEMICAL ENGINEERING TRANSACTIONS, vol. 43, p. 637-642, ISSN: 2283-9216, doi: 10.3303/CET1543107 3) Agrimi Gennaro, Pisano Isabella, Ricci Maria Antonietta, Palmieri Luigi (2015). 5. Microbial strain selection and development for the production of second-generation bioethanol. In: Gennaro Agrimi Isabella Pisano Maria Antonietta Ricci Luigi Palmieri. (a cura di): Michele Aresta Angela Dibenedetto Franck Dumeignil, BIOREFINERIES – An Introduction. p. 109-140, de Gruyter, ISBN: 9783110331585, doi: 10.1515/9783110331585-009 4) Ricci MA, Russo A, Pisano I, Palmieri L, de Angelis M, Agrimi G (2015). Improved 1,3-Propanediol synthesis from glycerol by the robust Lactobacillus reuteri strain DSM 20016. JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, vol. 25, p. 893-902, ISSN: 1017-7825, doi: dx.doi.org/10.4014/jmb.1411.11078 5) AGRIMI G, RICCI Maria Antonietta, PALMIERI Luigi, PISANO Isabella, VITALE Paola, PERNA Filippo Maria, SCILIMATI Antonio (2014). PROCEDIMENTO PER LA SINTESI DI ALCOL CHIRALI MEDIANTE IMPIEGO DI LACTOBACILLUS REUTERI. Domanda di brevetto italiano MI2014A002079, Università degli Studi di Bari 6) AGRIMI G, Annamaria Russo, Maria Antonietta Ricci (2014). Role of Mitochondrial Carriers in Metabolic Engineering. JOURNAL OF BIOPROCESSING & BIOTECHNIQUES, vol. 4, p. 1-2, ISSN: 2155-9821, doi: 10.4172/2155-9821.1000164 7) Agrimi G, Mena M, Izumi K, Pisano I, Germinario L, Fukuzaki H, Palmieri L, Blank L, Kitagaki H. (2014). Improved sake metabolic profile during fermentation due to increased mitochondrial pyruvate dissimilation.. FEMS YEAST RESEARCH, vol. 14, p. 249-260, ISSN: 1567-1356, doi: 10.1111/1567-1364.12120 8) Agrimi G, Pisano I, Palmieri L (2012). Process development and metabolic engineering for bioethanol production from lignocellulosic biomass. In: Biorefinery. p. 207-230, BERLINO: de Gruyter, ISBN: 978-3-11026028-1, doi: 10.1515/9783110260281.207 9) Marobbio CM, Pisano I, Porcelli V, Lasorsa FM, Palmieri L. Rapamycin reduces oxidative stress in frataxindeficient yeast cells. Mitochondrion. 2012 Jan;12(1):156-61. doi:10.1016/j.mito.2011.07.001. 10) Regalado A, Pierri CL, Bitetto M, Laera VL, Pimentel C, Francisco R, Passarinho J, Chaves MM, Agrimi G. Characterization of mitochondrial dicarboxylate/tricarboxylate transporters from grape berries. Planta. 2013 Mar;237(3):693-703. doi: 10.1007/s00425-012-1786-8. SUPPLEMENTARY MATERIAL Position of the components of the Research Groups Name Surname Position * Affiliation Gennaro Agrimi PA University of Bari Luigi Palmieri PO University of Bari Isabella Pisano RU University of Bari *: PO = Full professor; PA = Associate professor; RU = University researcher; CO = contract; PoD = Postdoctoral fellows; RC = CNR staff or other Institutions Research; T = technician, VR = visiting researcher, S = student Equipment Type Producer Year of acquisition Biostat B plus fermentation system Sartorius 2012 UPLC Acquity+ triple quadrupole mass spectrometer ESI Quattro Premier Waters 2004 Fast Real-Time PCR system 9700 HT Applied Biosystems 2009 GC-MS/MS Quattro Micro Waters - Agilent 2004 HPLC Alliance e2695 Waters 2002 Technical skills Biotransformations Metabolic flux analysis Gene cloning Expression and purification of recombinant proteins Metabolic engineering of microorganisms Fermentative process engineering Gene expression studies Enzimology