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Evaluating Pulses and Modified Fed-batch Feeding of Methanol to Increase Expression Level of Human Insulin Precursor in Pichia Pastoris High-Density Cultivation

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@article{IJASEIT15109,
   author = {Eko Wahyu Putro and Dini Nurdiani and - Hariyatun and Nuruliawaty Utami and Wien Kusharyoto and Ario Betha Juanssilfero and Rozy Ayu Silvia},
   title = {Evaluating Pulses and Modified Fed-batch Feeding of Methanol to Increase Expression Level of Human Insulin Precursor in Pichia Pastoris High-Density Cultivation},
   journal = {International Journal on Advanced Science, Engineering and Information Technology},
   volume = {12},
   number = {3},
   year = {2022},
   pages = {1001--1007},
   keywords = {Diabetes; human insulin precursor; Pichia pastoris; bioreactor; fed-batch.},
   abstract = {Production of sufficient insulin at a more affordable price is necessary. The increase in the number of people living with diabetes puts more burden on healthcare and the economy. P. pastoris is a promising host to produce human insulin precursors at a high yield in minimal medium and secretes low levels of endogenous protein impurities. Production of the precursor involves several parameters, including glycerol concentration, culture density, methanol concentration, and medium composition. This study evaluated the effect of those parameters on protein expression in the flask culture. Subsequently, fermentation in the bioreactor was carried on according to the information obtained from flask culture. Methanol feeding to induce protein expression was undertaken by pulses and fed-batch modes. The fed-batch method was modified from a standard technique by incorporating constant flow rates with variable feed concentrations. Cell density was determined based on optical density measurement at 600 nm and dry cell weight. Tricine SDS-PAGE and reversed-phase HPLC conducted protein analysis. The pulse feeding produced higher precursor concentrations at ~445 mg/L than modified fed-batch feeding at ~267 mg/L. However, the modified fed-batch feeding can be an alternative to producing human insulin precursors when a standard fed-batch feeding with variable flow rates and 100% (v/v) methanol feed is difficult to apply.},
   issn = {2088-5334},
   publisher = {INSIGHT - Indonesian Society for Knowledge and Human Development},
   url = {http://ijaseit.insightsociety.org/index.php?option=com_content&view=article&id=9&Itemid=1&article_id=15109},
   doi = {10.18517/ijaseit.12.3.15109}
}

EndNote

%A Putro, Eko Wahyu
%A Nurdiani, Dini
%A Hariyatun, -
%A Utami, Nuruliawaty
%A Kusharyoto, Wien
%A Juanssilfero, Ario Betha
%A Silvia, Rozy Ayu
%D 2022
%T Evaluating Pulses and Modified Fed-batch Feeding of Methanol to Increase Expression Level of Human Insulin Precursor in Pichia Pastoris High-Density Cultivation
%B 2022
%9 Diabetes; human insulin precursor; Pichia pastoris; bioreactor; fed-batch.
%! Evaluating Pulses and Modified Fed-batch Feeding of Methanol to Increase Expression Level of Human Insulin Precursor in Pichia Pastoris High-Density Cultivation
%K Diabetes; human insulin precursor; Pichia pastoris; bioreactor; fed-batch.
%X Production of sufficient insulin at a more affordable price is necessary. The increase in the number of people living with diabetes puts more burden on healthcare and the economy. P. pastoris is a promising host to produce human insulin precursors at a high yield in minimal medium and secretes low levels of endogenous protein impurities. Production of the precursor involves several parameters, including glycerol concentration, culture density, methanol concentration, and medium composition. This study evaluated the effect of those parameters on protein expression in the flask culture. Subsequently, fermentation in the bioreactor was carried on according to the information obtained from flask culture. Methanol feeding to induce protein expression was undertaken by pulses and fed-batch modes. The fed-batch method was modified from a standard technique by incorporating constant flow rates with variable feed concentrations. Cell density was determined based on optical density measurement at 600 nm and dry cell weight. Tricine SDS-PAGE and reversed-phase HPLC conducted protein analysis. The pulse feeding produced higher precursor concentrations at ~445 mg/L than modified fed-batch feeding at ~267 mg/L. However, the modified fed-batch feeding can be an alternative to producing human insulin precursors when a standard fed-batch feeding with variable flow rates and 100% (v/v) methanol feed is difficult to apply.
%U http://ijaseit.insightsociety.org/index.php?option=com_content&view=article&id=9&Itemid=1&article_id=15109
%R doi:10.18517/ijaseit.12.3.15109
%J International Journal on Advanced Science, Engineering and Information Technology
%V 12
%N 3
%@ 2088-5334

IEEE

Eko Wahyu Putro,Dini Nurdiani,- Hariyatun,Nuruliawaty Utami,Wien Kusharyoto,Ario Betha Juanssilfero and Rozy Ayu Silvia,"Evaluating Pulses and Modified Fed-batch Feeding of Methanol to Increase Expression Level of Human Insulin Precursor in Pichia Pastoris High-Density Cultivation," International Journal on Advanced Science, Engineering and Information Technology, vol. 12, no. 3, pp. 1001-1007, 2022. [Online]. Available: http://dx.doi.org/10.18517/ijaseit.12.3.15109.

RefMan/ProCite (RIS)

TY  - JOUR
AU  - Putro, Eko Wahyu
AU  - Nurdiani, Dini
AU  - Hariyatun, -
AU  - Utami, Nuruliawaty
AU  - Kusharyoto, Wien
AU  - Juanssilfero, Ario Betha
AU  - Silvia, Rozy Ayu
PY  - 2022
TI  - Evaluating Pulses and Modified Fed-batch Feeding of Methanol to Increase Expression Level of Human Insulin Precursor in Pichia Pastoris High-Density Cultivation
JF  - International Journal on Advanced Science, Engineering and Information Technology; Vol. 12 (2022) No. 3
Y2  - 2022
SP  - 1001
EP  - 1007
SN  - 2088-5334
PB  - INSIGHT - Indonesian Society for Knowledge and Human Development
KW  - Diabetes; human insulin precursor; Pichia pastoris; bioreactor; fed-batch.
N2  - Production of sufficient insulin at a more affordable price is necessary. The increase in the number of people living with diabetes puts more burden on healthcare and the economy. P. pastoris is a promising host to produce human insulin precursors at a high yield in minimal medium and secretes low levels of endogenous protein impurities. Production of the precursor involves several parameters, including glycerol concentration, culture density, methanol concentration, and medium composition. This study evaluated the effect of those parameters on protein expression in the flask culture. Subsequently, fermentation in the bioreactor was carried on according to the information obtained from flask culture. Methanol feeding to induce protein expression was undertaken by pulses and fed-batch modes. The fed-batch method was modified from a standard technique by incorporating constant flow rates with variable feed concentrations. Cell density was determined based on optical density measurement at 600 nm and dry cell weight. Tricine SDS-PAGE and reversed-phase HPLC conducted protein analysis. The pulse feeding produced higher precursor concentrations at ~445 mg/L than modified fed-batch feeding at ~267 mg/L. However, the modified fed-batch feeding can be an alternative to producing human insulin precursors when a standard fed-batch feeding with variable flow rates and 100% (v/v) methanol feed is difficult to apply.
UR  - http://ijaseit.insightsociety.org/index.php?option=com_content&view=article&id=9&Itemid=1&article_id=15109
DO  - 10.18517/ijaseit.12.3.15109

RefWorks

RT Journal Article
ID 15109
A1 Putro, Eko Wahyu
A1 Nurdiani, Dini
A1 Hariyatun, -
A1 Utami, Nuruliawaty
A1 Kusharyoto, Wien
A1 Juanssilfero, Ario Betha
A1 Silvia, Rozy Ayu
T1 Evaluating Pulses and Modified Fed-batch Feeding of Methanol to Increase Expression Level of Human Insulin Precursor in Pichia Pastoris High-Density Cultivation
JF International Journal on Advanced Science, Engineering and Information Technology
VO 12
IS 3
YR 2022
SP 1001
OP 1007
SN 2088-5334
PB INSIGHT - Indonesian Society for Knowledge and Human Development
K1 Diabetes; human insulin precursor; Pichia pastoris; bioreactor; fed-batch.
AB Production of sufficient insulin at a more affordable price is necessary. The increase in the number of people living with diabetes puts more burden on healthcare and the economy. P. pastoris is a promising host to produce human insulin precursors at a high yield in minimal medium and secretes low levels of endogenous protein impurities. Production of the precursor involves several parameters, including glycerol concentration, culture density, methanol concentration, and medium composition. This study evaluated the effect of those parameters on protein expression in the flask culture. Subsequently, fermentation in the bioreactor was carried on according to the information obtained from flask culture. Methanol feeding to induce protein expression was undertaken by pulses and fed-batch modes. The fed-batch method was modified from a standard technique by incorporating constant flow rates with variable feed concentrations. Cell density was determined based on optical density measurement at 600 nm and dry cell weight. Tricine SDS-PAGE and reversed-phase HPLC conducted protein analysis. The pulse feeding produced higher precursor concentrations at ~445 mg/L than modified fed-batch feeding at ~267 mg/L. However, the modified fed-batch feeding can be an alternative to producing human insulin precursors when a standard fed-batch feeding with variable flow rates and 100% (v/v) methanol feed is difficult to apply.
LK http://ijaseit.insightsociety.org/index.php?option=com_content&view=article&id=9&Itemid=1&article_id=15109
DO  - 10.18517/ijaseit.12.3.15109