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Serin alkali proteaz üretimi için biyoproses tasarım parametrelerinin araştırılması

Bioprocesses design parameters for serine alkaline protease production

  1. Tez No: 35023
  2. Yazar: PINAR ÇALIK
  3. Danışmanlar: PROF. DR. H. TUNÇER ÖZDAMAR
  4. Tez Türü: Yüksek Lisans
  5. Konular: Kimya Mühendisliği, Chemical Engineering
  6. Anahtar Kelimeler: Belirtilmemiş.
  7. Yıl: 1994
  8. Dil: Türkçe
  9. Üniversite: Ankara Üniversitesi
  10. Enstitü: Fen Bilimleri Enstitüsü
  11. Ana Bilim Dalı: Kimya Mühendisliği Ana Bilim Dalı
  12. Bilim Dalı: Belirtilmemiş.
  13. Sayfa Sayısı: 151

Özet

The bioprocess design parameters for serine alkaline protease (SAP) production by using a mutant of Bacillus subtilis was investiga ted; and a method for quantitative protein analysis were developed by using HPCE. Effects of pH and temperature on the bioprocess were in vestigated simultaneously with a programme at T»37,45,50,55°C and at e^ery temperature least with seven pH values in the range of pH«5-12; and the optimum values were determined as pH=10 and 55°C. Because the culture conditions that promote production of SAP and neutral prote- ase(NP) were found significantly different from each bother and from the culture conditions promoting cell growth; it was therefore essen tial to design a media in order to maintain a high cell density cultu re under conditions promoting SAP production rather then NP. Since the components of the medium regulates the SAP activity and concentration, starting from the semi -defined media in the literature a set of compo nents and their effects on the design af the media were investigated. Inorganic compounds Ca, Mg+Z and Cu+Z were increased the protease activity and SAP concentration; however Zn+Z has a negative effect. 0.25g/dnr was found as the optimum for L-serine; nevertheless, it has not much effect on enzyme activity in the defined media. It was found that fructose increased the activity and the stability of SAP; however it has an inhibitor effect on the production of NP. While pepton was increasing the concentration of SAP, yeast extract increased NP con centration. In the fifth phase of the programme, by using two alter native mediums with and without fructose the course of activity, SAP and NP concentrations were investigated. After 96h of operation, the fructose containing and the fructose free mediums produced 1.63/gdnr and 1.59g/dnr SAP, respectively. However, the NP productions were found none in the first, but 3.2g/dm3 in the latter. In the last phase, by using the best process conditions found, in a temperature, pH, foam and oxygen controlled pilot-scale reactor the results were tested during with least 50 hours of runs. KEY WORDS: Serine alkaline protease, neutral protease, production of protease, capiller electroforesis, design parametres, Bad 7 lus subtnis. İV

Özet (Çeviri)

The bioprocess design parameters for serine alkaline protease (SAP) production by using a mutant of Bacillus subtilis was investiga ted; and a method for quantitative protein analysis were developed by using HPCE. Effects of pH and temperature on the bioprocess were in vestigated simultaneously with a programme at T»37,45,50,55°C and at e^ery temperature least with seven pH values in the range of pH«5-12; and the optimum values were determined as pH=10 and 55°C. Because the culture conditions that promote production of SAP and neutral prote- ase(NP) were found significantly different from each bother and from the culture conditions promoting cell growth; it was therefore essen tial to design a media in order to maintain a high cell density cultu re under conditions promoting SAP production rather then NP. Since the components of the medium regulates the SAP activity and concentration, starting from the semi -defined media in the literature a set of compo nents and their effects on the design af the media were investigated. Inorganic compounds Ca, Mg+Z and Cu+Z were increased the protease activity and SAP concentration; however Zn+Z has a negative effect. 0.25g/dnr was found as the optimum for L-serine; nevertheless, it has not much effect on enzyme activity in the defined media. It was found that fructose increased the activity and the stability of SAP; however it has an inhibitor effect on the production of NP. While pepton was increasing the concentration of SAP, yeast extract increased NP con centration. In the fifth phase of the programme, by using two alter native mediums with and without fructose the course of activity, SAP and NP concentrations were investigated. After 96h of operation, the fructose containing and the fructose free mediums produced 1.63/gdnr and 1.59g/dnr SAP, respectively. However, the NP productions were found none in the first, but 3.2g/dm3 in the latter. In the last phase, by using the best process conditions found, in a temperature, pH, foam and oxygen controlled pilot-scale reactor the results were tested during with least 50 hours of runs. KEY WORDS: Serine alkaline protease, neutral protease, production of protease, capiller electroforesis, design parametres, Bad 7 lus subtnis. İV

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