Path: TopS1-Final ProjectMicrobiology-SITH2017

PENGEMBANGAN KULTUR DENSITAS TINGGI Pichia pastoris GS115 TERINTEGRASI 4 SALINAN KASET sHBsAg

The Development of High Density Culture of Pichia pastoris GS115 Integrated with 4 coppy of sHBsAg Cassettes

Undergraduate Theses from JBPTITBPP / 2017-09-27 10:14:14
By : LIM EDRIA KOSASIH (NIM : 10413015), School of Life Sciences and Technology
Created : 2017-09-06, with 1 files

Keyword : ekstrak ragi, kultivasi curah umpan, oksigen, Pichia pastoris, sHBsAg

Penelitian terdahulu telah berhasil mengembangkan Pichia pastoris rekombinan








GS115 yang mengandung 4 Salinan Kaset sHBsAg sebagai sistem ekspresi protein








sHBsAg. Protein sHBsAg merupakan bahan baku vaksin Hepatitis B yang hingga








saat ini masih diimpor. Penelitian selanjutnya menunjukan bahwa penggunaan








komposisi medium pertumbuhan yang disarankan oleh berbagai penelitian tidak








dapat memberikan konsentrasi biomassa yang diinginkan. Dalam penelitian ini,








dilakukan modifikasi komposisi medium dan teknik kultivasi curah untuk








menumbuhkan Pichia pastoris GS115 agar dapat menghasilkan kultur dengan








konsentrasi biomassa yang tinggi. Hasil penelitian menunjukan bahwa penambahan








ekstrak ragi dengan konsentrasi 6,89 g/l ke dalam medium mampu menghasilkan








kultur dengan biomassa Pichia pastoris GS115 hingga 38,75 g/l. Konsentrasi








biomassa ini lebih tinggi dibandingkan dengan konsentrasi biomassa pada kultur








yang tidak mendapatkan penambahan ekstrak ragi, yang hanya mencapai kepadatan








16,98 g/l. Penelitian selanjutnya menunjukan bahwa penerapan metode kultivasi








curah umpan dapat meningkatkan konsentrasi biomassa Pichia pastoris GS115








pada kultur lebih tinggi lagi dan pasokan oksigen pada kultur sangat mempengaruhi








biomassa yang diperoleh. Hasil penelitian ini menunjukan bahwa kultur curah








umpan yang diberi pasokan 4 liter per menit (LPM) udara dan 0,26 LPM oksigen








menghasilkan kultur dengan kepadatan yang paling tinggi, yaitu 119,47 g/l, diikuti








dengan kultur yang diberi pasokan 4 LPM udara dan 0,58 LPM oksigen, yaitu








111,52 g/l, pasokan 4 LPM udara dan 1,2 LPM oksigen, yaitu 95,78 g/l dan 4 LPM








udara tanpa penambahan oksigen, yaitu 39,55 g/l. Jadi, penambahan ektrak ragi dan








pasokan oksigen dapat meningkatkan konsentrasi biomassa Pichia pastoris GS115.

Description Alternative :

The latest research has successfully developed recombinant Pichia pastoris GS115








which contain 4 cassettes of sHBsAg as a sHBsAg protein expression system. Until








recently, Indodesia still imports sHBsAg protein which is the main material in








Hepatitis B vaccine production. Further research shows that media which has been








suggested in research papers is unable to produce high concentration of biomass.








In this research, modification of media composition and fed-batch cultivation








technique are conducted to achieve high concentration of biomass. The result








shows that the addition of 6.89 g/L yeast extract to the cultivation media was able








to produce 38.75 g/L Pichia pastoris GS115 biomass. This number is higher than








the biomass concentration obtained by cultivation without the addition of yeast








extract which was 16.98 g/L. Further research proves that fed-batch cultivation is








able to increase biomass concentration of Pichia pastoris GS115. Moreover,








oxygen supply affects obtained biomass. Fed-batch cultivation with 4 liter per








minute (LPM) of air and 0.26 LPM oxygen is able to achieve the highest culture








density which was 119.47 g/L, followed by culture supplied with 4 LPM of air and








0.58 LPM oxygen which was 111.52 g/L, 4 LPM of air and 1.2 LPM oxygen which








was 95.78 g/L, and 4 LPM of air whithout oxygen supply which was 39.55 g/L. In








conclusion, the addition of yeast extract and oxygen supply is able to increase








biomass concentration of Pichia pastoris GS115.

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