TY - JOUR
T1 - Isolation of cDNA from Jacaratia mexicana encoding a mexicain-like cysteine protease gene
AU - Ramos-Martínez, Erick M.
AU - Herrera-Ramírez, Alejandra C.
AU - Badillo-Corona, Jesús Agustín
AU - Garibay-Orijel, Claudio
AU - González-Rábade, Nuria
AU - Oliver-Salvador, María del Carmen
N1 - Funding Information:
Research in the laboratory of MCOS was supported by Instituto Politécnico Nacional-SIP ( 20100222 and 20110944 ). JABC, MCOS and CGO are COFAA and EDI-IPN scholars. The authors also want to thank Professor Roberto Briones-Martínez for providing the J. mexicana seeds used in this research.
PY - 2012/7/1
Y1 - 2012/7/1
N2 - Cysteine proteases (CPs) from the C1 family, which are similar to papain, can be found in animals and plants, as well as some viruses and prokaryotes. These enzymes have diverse physiological functions and are thus very attractive for science and industry. Jacaratia mexicana, a member of the Caricaceae plant family, contains several CPs, the principal being mexicain, found to favorably compete against papain for many industrial applications due to its high stability and specific activity. In this study, leaves of J. mexicana were used to isolate a CP-coding gene, similar to those that code for mexicain and chymomexicain. By using rapid amplification of cDNA ends (RACE) as well as oligonucleotide design from papain-like conserved amino acids (aa), a sequence of 1404. bp consisting of a 5' terminal untranslated region (UTR) of 153. bp, a 3' terminal UTR of 131. bp, with a polyadenylation (poly(A)) signal sequence and a poly(A) tail, and an open reading frame (ORF) of 1046. bp, was obtained by overlapping three partial sequences. Two full-length cDNA sequences that encode for mexicain-like proteases were cloned from mRNA (JmCP4 and JmCP5). JmCP4 is predicted to have an ORF of 1044. bp, which codifies for polypeptides that have a 26 aa signal peptide region, a 108 aa propeptide region and a mature enzyme of 214 aa. A 969. bp fragment (JmCP5) encodes for a partial sequence of a CP gene, without the signal peptide region but with a full-length propeptide region. The sequence analysis showed that this protease presented a high similarity to other plant CPs from J. mexicana, Vasconcellea cundinamarcensis, Vasconcellea stipulata, and Carica papaya, among others, mainly at the conserved catalytic site. Obtaining the sequence of this CP gene from J. mexicana provides an alternative for production in a standard system and could be an initial step towards the commercialization of this enzyme.
AB - Cysteine proteases (CPs) from the C1 family, which are similar to papain, can be found in animals and plants, as well as some viruses and prokaryotes. These enzymes have diverse physiological functions and are thus very attractive for science and industry. Jacaratia mexicana, a member of the Caricaceae plant family, contains several CPs, the principal being mexicain, found to favorably compete against papain for many industrial applications due to its high stability and specific activity. In this study, leaves of J. mexicana were used to isolate a CP-coding gene, similar to those that code for mexicain and chymomexicain. By using rapid amplification of cDNA ends (RACE) as well as oligonucleotide design from papain-like conserved amino acids (aa), a sequence of 1404. bp consisting of a 5' terminal untranslated region (UTR) of 153. bp, a 3' terminal UTR of 131. bp, with a polyadenylation (poly(A)) signal sequence and a poly(A) tail, and an open reading frame (ORF) of 1046. bp, was obtained by overlapping three partial sequences. Two full-length cDNA sequences that encode for mexicain-like proteases were cloned from mRNA (JmCP4 and JmCP5). JmCP4 is predicted to have an ORF of 1044. bp, which codifies for polypeptides that have a 26 aa signal peptide region, a 108 aa propeptide region and a mature enzyme of 214 aa. A 969. bp fragment (JmCP5) encodes for a partial sequence of a CP gene, without the signal peptide region but with a full-length propeptide region. The sequence analysis showed that this protease presented a high similarity to other plant CPs from J. mexicana, Vasconcellea cundinamarcensis, Vasconcellea stipulata, and Carica papaya, among others, mainly at the conserved catalytic site. Obtaining the sequence of this CP gene from J. mexicana provides an alternative for production in a standard system and could be an initial step towards the commercialization of this enzyme.
KW - Cysteine protease
KW - Jacaratia mexicana
KW - Mexicain
KW - Papain
UR - http://www.scopus.com/inward/record.url?scp=84860914837&partnerID=8YFLogxK
U2 - 10.1016/j.gene.2012.04.018
DO - 10.1016/j.gene.2012.04.018
M3 - Artículo
C2 - 22543019
SN - 0378-1119
VL - 502
SP - 60
EP - 68
JO - Gene
JF - Gene
IS - 1
ER -