Resúmenes

Material suplementario S1
Citación:
Villena G, Hernández- Macedo M, Samolski I. 2020. Memorias Del Primer Congreso
Internacional de Biotecnología e Innovación - ICBi 2018. Material suplementario S1.
Revista Peruana De Biología 27 (1), 5-14. https://doi.org/10.15381/rpb.v27i1.17624

Trabajos presentados al I Congreso
Internacional de Biotecnología e
innovación (ICBi), 9 - 12 de julio de
2018, Universidad Nacional Agraria
La Molina, Lima, Perú.
Editoras:
Ilanit Samolski Klein
Maria Lucila Hernández-Macedo
Gretty Katherina Villena Chávez
Resúmenes del I Congreso Internacional de Biotecnología e innovación
Contribution
Section II: Industrial and Environmetal Biotechnology
Potential application of commercial enzyme preparations for the produc-
tion of isomaltooligosaccharides from maltose
Berrocal, RC.1* Vega, PR1,2; Chico, LH3; Carranza, C.1
1 Universidad Nacional Mayor de San Marcos, Laboratorio del Grupo de Investigación de Procesos
Biotecnológicos y Alimentarios, Lima, Perú. - 2 Universidad Nacional Mayor de San Marcos, Depar-
tamento de Bioquímica, Facultad de Farmacia y Bioquímica, Lima, Perú. - 3 Universidad Nacional del
Santa, Laboratorio de Fisicoquímica, Departamento de Agroindustria, Facultad de Ingeniería, Nuevo
Chimbote, Perú.
* Corina.berrocal11@gmail.com
Keywords: transglycosylation activity, hydrolysis activity, ?-glucosidase, panose
Isomaltooligosaccharides (IMOs) are glucose oligomers containing one or more ?-(1,6) glycosidic bonds, although
they are marketed as a mixture of glucosyl saccharides with both ?-(1,6) and ?-(1,4) bondsIMOs are emerging
prebiotics and exhibit unique physicochemical properties for the food industryThey are produced from maltose
by means of ?-glucosidases (EC 3.2.1.20) transglycosylation reactionsHowever, few ?-glucosidases exhibit the
required level of transglycosylation for industrial application because several of these enzymes catalyze the maltose
hydrolysis instead of the synthesis of IMOsTherefore, we performed a screening of available commercial enzyme
preparations for the potential production of IMOs from maltose and determined the effect of temperature on
the transglycosylation and hydrolysis activities of the selected enzyme preparationUsing maltose as substrate,
the activities were determined by the initial reaction rates of panose formation and glucose releasedThe reaction
mixture consisted of 1.9 mL of 300 g/L maltose in 50 mM sodium acetate buffer at pH 5.0 and 0.1 mL of either
enzyme or diluted enzyme using the same bufferThe mixture was incubated at 50 °C and stirred at 150 rpmThe
samples were analyzed for their carbohydrate composition by high-performance liquid chromatography (HPLC)
The effect of temperature on the activities was studied in the range from 35 to 60 °C while the other reaction
conditions were constant as the previously mentioned valuesThe experiments were performed in duplicateThe
enzyme preparation called Cellulase DS was selected from six enzyme preparations, since it exhibited the highest
transglycosylation activity (464 ± 7.1 UI/g) and ratio of the synthesis and hydrolysis activities (1.1 ± 0.06)In addition,
this enzyme is highly selective for the panose synthesis compared to the enzyme called Transglucosidase L, which
is used on an industrial scaleAs expected, the transglycosylation and hydrolysis activities significantly increased as
the temperature increased, but the hydrolysis was more sensitive to temperature than the synthesisThus, the ratio
of the synthesis and hydrolysis activities was higher at 35 °C (R =1.25 ± 0.039)The results indicate that there are
commercial enzyme preparations exhibiting a secondary transglycosylation activity, which can serve as a source of
?-glucosidase for the enzymatic production of IMOs from maltose.
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Contribution
Section II: Industrial and Environmetal Biotechnology
Proteomic analysis of the cyanotrophic strain Alcaligenes aquatilis after its
exposure to cyanide by MALDI TOF / TOF.
Cubas, C1* Liza, V1; Cornejo, M2; Mialhe, E3
1Universidad Nacional de Tumbes (UNT), Laboratorio de Biología Molecular de la Facultad de Inge-
niería pesquera, Tumbes, Perú. - 2Cooperativa de Trabajadores BIOTECOOP, Departamento de Biotec-
nología Ambiental. - 3INCA’BIOTEC S.A.C, Laboratorio de Biotecnología Molecular, Tumbes, Perú
* carloscubaszuniga92@gmail.com
Keywords: Cyanide, bioremediation, nitrilase, exoproteome, enzyme
Cyanide leaching is the main process used in the mining industry for the extraction of gold and silver in Peru and
worldwideCyanide is a highly toxic and harmful compound to the environment; however, there are microorganisms
with the ability to degrade this compoundThe cyanotrophic strain Alcaligenes aquatilis isolated from mining areas
affected by cyanide in the La Libertad region, was studied due to its high performance to resist and degrade cyanideIn
order to examine the physiological responses of cyanide degradation by this bacterium, a bottom-up proteomics
approach by proteolytic digestion of proteins prior to analysis by MALDI TOF / TOF mass spectrometry was applied, in
addition a genomic identification of cyanide degrading enzymes by endpoint PCR was doneFor the proteomic study
Alcaligenes aquatilis was cultured in a minimal medium with 200 ppm of sodium cyanide (NaCN), the intracellular
and extracellular proteins were extracted and migrated in SDS-PAGE gel for separation by molecular weight, the
differential bands were cut, processed and analyzed in the MALDI TOF / TOFFrom the genomic DNA extracted from
this microorganism, the endpoint PCR was carried out with primers target to the NHase gene (nitrilase)Alcaligenes
aquatilis cultivated in minimal medium with cyanide as the sole source of nitrogen, managed to degrade it in 24
hoursOn the other hand, enzymes involved in the cyanide degradation, such as cyanase, nitrilasa and cyanate
hydratase were identified by MALDI TOF / TOF; moreover, transmembrane proteins involved in the transport of
cyanide and cyanide compounds as MFS cyanate transporters and ABC cyanate transporters were identified in the
intracellular samplesIn addition, proteins such as the siderophore synthetase that participate in the biosynthesis
of siderophores were identified, these are key for the resistance of the strain to cyanideThe transcription regulator
CynR, also identified, fulfills the function of modulating the expression of the enzyme cyanase in the presence of
cyanideBesides, the nitrilase enzyme was found in the exoproteome samplesAt the genomic level, amplification of
the NHase (nitrilase) gene was achieved by PCRThis work shows that a novel cyanotrophic bacteria has the potential
to be used for bioremediation based on its genomic and proteomic characteristics.
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Material suplementario S1, Rev. peru. biol. 27(1): 21 - 053 (March 2020) S1-21