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