12055

11044

كارشناسي ارشد

بيوتكنولوژي

اصفهان: دانشگاه صنعتي اصفهان، دانشكده كشاورزي

1395

دوازده، 78ص.: مصور، جدول، نمودار

ص. ع. به فارسي و انگليسي

آذر شاه پيري

اصغر طاهري كفراني

مجيد طالبي، اميرحسين مهدوي

1395/11/10

كتابنامه

كشاورزي

مهندسي كشاورزي

ID11044

Study of Digestibility of Milk by Using Recombinant Systems of Rice Thioredoxin Thioredoxin Reductase Tahere Shahriari Farfani Tahereshahriari2015@gmail com Date of submission January 1 2017 Department of Agricultural Biotehnology Isfahan University of Technology Isfahan 84156 83111 Iran Degere M Sc Language FarsiSupervisors A shahpiri Shahpiri715@yahoo com A Taheri Kafrani A taheri ast ui ac irAbstract lactoglobulin BLG is a member of lipocalin superfamily of transporters for small hydrophobic moleculessuch as retinoids fatty acids drugs and vitamins BLG structure has 2 intramolecular disulfide bonds and onefree cystein in cys121 BLG is one of the major bovine milk allergen that disulfide bonds are responsible forbovine milk allergy and low digestibility It seems that the digestibility of BLG will be increased via thereduction of disulfide bonds The reducing agents such as DTT is able to reduce disulfide bonds and increase thethe BLG digestibility However due to the toxic effect of DTT the use of this agent is not applicable Therefore this can be substituted by natural enzymes In the present study we aim to study the effect of plant thioredoxinsystems on the digestibility of milk BLG Thioredoxins Trxs are small and abundant disulfide reductase in allorganisms Trxs are characterized with WC G P PC in active site Trxs are reduced themselves with NADPHvia NADPH dependent thioredoxin reductase NTR Previously the genes encoding three rice Trx OsTrx1 OsTrx20 and OsTrx23 and the gene encoding OsNTR1were cloned and heterologously expressed in E coli Inthis study these proteins were produced and purified in considerable amounts The activity of Trxs wasconfirmed by insulin assay Moreover the interaction of OsNTR1 with each of OsTrxs was performed usingDTNB assay The result of this assay showed that OsNTR1 was able to reduce OsTrx1 and OsTrx23 However OsTrx20 was not reduced by OsNTR1 Instead this isoform was reduced by glutathione GSH The isoformOsTrx1 can also be reduced by GSH Therefore in this study the effect of four systems OsNTR1 OsTrx23 OsNTR1 OsTrx1 GSH OsTrx20 GSH OsTrx1 on the digestibility of BLG was studied To this end BLG waspre treated by these systems at 4 25 37 and 55 C and then digested with trypsin The digestability was studiedusing SDS PAGE and HPLC The results revealed that different Trx system significantly affect on the BLGdigestability However there was no significant difference between the effect of different systems In addition the results show that Trx systems work in 4 25 and 37 C Nevertheless these systems were not affected at 55 Cwhich could be due to the unstability of NTR in this temperature Keywords NTR Trx system glutathione beta lactoglobulin digestibility trypsin reversephase liquid chromatography

آذر شاه پيري

اصغر طاهري كفراني

مجيد طالبي، اميرحسين مهدوي