High-Level Expression of Recombinant Bovine Lactoferrin in Pichia pastoris with Antimicrobial Activity.
In this study, bovine lactoferrin (bLf), an iron-binding glycoprotein considered an important nutraceutical protein because of its several properties, was expressed in Pichia pastoris KM71-H under AOX1 promoter control, using pJ902 as the recombinant plasmid. Dot blotting analysis revealed the expression of recombinant bovine lactoferrin (rbLf) in Pichia pastoris. After Bach fermentation and purification by molecular exclusion, we obtained an expression yield of 3.5 g/L of rbLf. rbLf and predominantly pepsin-digested rbLf (rbLfcin) demonstrated antibacterial activity against Escherichia coli (E. coli) BL21DE3, Staphylococcus aureus (S. aureus) FRI137, and, in a smaller percentage, Pseudomonas aeruginosa (Ps. Aeruginosa) ATCC 27833. The successful expression and characterization of functional rbLf expressed in Pichia pastoris opens a prospect for the development of natural antimicrobial agents produced recombinantly.
Laboratorio de Biotecnolog&#237;a 1, Facultad de Ciencias Qu&#237;micas, Universidad Aut&#243;noma de Chihuahua, Circuito 1, Nuevo Campus Universitario, Chihuahua CP 31125, Mexico. blancaflor_igf@hotmail.com.
Blanca
B
Iglesias-Figueroa
Laboratorio de Biotecnolog&#237;a 1, Facultad de Ciencias Qu&#237;micas, Universidad Aut&#243;noma de Chihuahua, Circuito 1, Nuevo Campus Universitario, Chihuahua CP 31125, Mexico. nvaldiviezogodina@gmail.com.
Norberto
N
Valdiviezo-Godina
Laboratorio de Biotecnolog&#237;a 1, Facultad de Ciencias Qu&#237;micas, Universidad Aut&#243;noma de Chihuahua, Circuito 1, Nuevo Campus Universitario, Chihuahua CP 31125, Mexico. tsiqueiros@uach.mx.
Tania
T
Siqueiros-Cendón
Laboratorio de Biotecnolog&#237;a, Campus de Ciencias Agropecuarias, Universidad Aut&#243;noma de Nuevo Le&#243;n, Francisco Villa S/N Col. Ex hacienda El Canad&#225;, General Escobedo, Nuevo Le&#243;n 66054, Mexico. ssinagawa@gmail.com.
Sugey
S
Sinagawa-García
Laboratorio de Biotecnolog&#237;a 1, Facultad de Ciencias Qu&#237;micas, Universidad Aut&#243;noma de Chihuahua, Circuito 1, Nuevo Campus Universitario, Chihuahua CP 31125, Mexico. sareval@uach.mx.
Sigifredo
S
Arévalo-Gallegos
Laboratorio de Biotecnolog&#237;a 1, Facultad de Ciencias Qu&#237;micas, Universidad Aut&#243;noma de Chihuahua, Circuito 1, Nuevo Campus Universitario, Chihuahua CP 31125, Mexico. qrascon@uach.mx.
Quintín
Q
Rascón-Cruz
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002417:Cattle; D005285:Fermentation; D007218:Industrial Microbiology / Q000379:methods; D007781:Lactoferrin / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D010843:Pichia / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D000900:Anti-Bacterial Agents; D011994:Recombinant Proteins; D007781:Lactoferrin
Pichia pastoris; antibacterial activity; lactoferricin; recombinant bovine lactoferrin