Resveratrol production in bioreactor: Assessment of cell physiological states and plasmid segregational stability

نویسندگان

  • Margarida S. Afonso
  • Susana Ferreira
  • Fernanda C. Domingues
  • Filomena Silva
چکیده

Resveratrol is a plant secondary metabolite commonly found in peanuts and grapevines with significant health benefits. Recombinant organisms can produce large amounts of resveratrol and, in this work, Escherichia coli BW27784 was used to produce resveratrol in bioreactors while monitoring cell physiology and plasmid stability through flow cytometry and real-time qPCR, respectively. Initially, the influence of culture conditions and precursor addition was evaluated in screening assays and the data gathered was used to perform the bioreactor assays, allowing the production of 160 μg/mL of resveratrol. Cellular physiology and plasmid instability affected the final resveratrol production, with lower viability and plasmid copy numbers associated with lower yields. In sum, this study describes new tools to monitor the bioprocess, evaluating the effect of culture conditions, and its correlation with cell physiology and plasmid segregational stability, in order to define a viable and scalable bioprocess to fulfill the need for larger quantities of resveratrol.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Evaluation of the hok/sok killer locus for enhanced plasmid stability.

The effectiveness of the hok/sok plasmid stability locus and mechanism of cloned-gene loss was evaluated in shake-flask cultures. Addition of the hok/sok locus dramitically increasedapparent plasmid segregational stability to the hok/sok(-) control. In terms of the number of generations before 10%of the population became plasmid-free, segregational stability was increased by 11- to 20-fold in d...

متن کامل

Physiological and Morphological Changes of Recombinant E. coli During Over-Expression of Human Interferon-g in HCDC

The objective of this research was to investigate the influence of the over-expression of recombinant interferon-g during high cell density cultivation on cellular characteristics of recombinant E. coli. Batch and fed-batch culture techniques were employed to grow Escherichia coli BL21 for production of human gamma-interferon in pET expression system. Final cell densities in batch and fed-batch...

متن کامل

A cell engineering strategy to enhance supercoiled plasmid DNA production for gene therapy

With the recent revival of the promise of plasmid DNA vectors in gene therapy, a novel synthetic biology approach was used to enhance the quantity, (yield), and quality of the plasmid DNA. Quality was measured by percentage supercoiling and supercoiling density, as well as improving segregational stability in fermentation. We examined the hypothesis that adding a Strong Gyrase binding Site (SGS...

متن کامل

Enhanced production of recombinant streptokinase in Escherichia coli using fed-batch culture.

Fed-batch culture strategy is often used for increasing production of heterologous recombinant proteins in Escherichia coli. This study was initiated to investigate the effects of dissolved oxygen concentration (DOC), complex nitrogen sources and pH control agents on cell growth and intracellular expression of streptokinase (SK) in recombinant E. coli BL21(DE3). Increase in DOC set point from 3...

متن کامل

Modelling of Bioprocess for Streptokinase Production Using Mechanistic and Neural Network Approaches

Streptokinase is a vital fibrinolytic drug produced by くhemolytic streptococci often used to treat myocardial infarction and pulmonary embolism. While growing recombinant strain of E.coli to produce streptokinase in a bioreactor, we are dealing with a controlled environment experiencing the role several indispensable factors that are associated to structured and unstructured aspects of the syst...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:

دوره 5  شماره 

صفحات  -

تاریخ انتشار 2015