fed batch culture
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Author(s):  
Yan Du ◽  
Fangyu Cheng ◽  
Miaomiao Wang ◽  
Chunmeng Xu ◽  
Huimin Yu

Hyaluronic acid (HA) is composed of alternating d-glucuronic acid and N-acetyl-d-glucosamine, with excellent biocompatibility and water retention capacity. To achieve heterologous biosynthesis of HA, Corynebacterium glutamicum, a safe GRAS (generally recognized as safe) host, was utilized and metabolically engineered previously. In this work, to achieve further enhancement of HA yield, four strategies were proposed and performed separately first, i.e., (1) improvement of glucose uptake via iolR gene knockout, releasing the inhibition of transporter IolT1/IolT2 and glucokinases; (2) intensification of cardiolipin synthesis through overexpression of genes pgsA1/pgsA2/cls involved in cardiolipin synthesis; (3) duly expressed Vitreoscilla hemoglobin in genome, enhancing HA titer coupled with more ATP and improved NAD+/NADH (>7.5) ratio; and (4) identification of the importance of glutamine for HA synthesis through transcriptome analyses and then enhancement of the HA titer via its supplement. After that, we combined different strategies together to further increase the HA titer. As a result, one of the optimal recombinant strains, Cg-dR-CLS, yielded 32 g/L of HA at 60 h in a fed-batch culture, which was increased by 30% compared with that of the starting strain. This high value of HA titer will enable the industrial production of HA via the engineered C. glutamicum.


2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Yohei Ishibashi ◽  
Hatsumi Goda ◽  
Rie Hamaguchi ◽  
Keishi Sakaguchi ◽  
Takayoshi Sekiguchi ◽  
...  

AbstractThe demand for n-3 long-chain polyunsaturated fatty acids (n-3LC-PUFAs), such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), will exceed their supply in the near future, and a sustainable source of n-3LC-PUFAs is needed. Thraustochytrids are marine protists characterized by anaerobic biosynthesis of DHA via polyunsaturated fatty acid synthase (PUFA-S). Analysis of a homemade draft genome database suggested that Parietichytrium sp. lacks PUFA-S but possesses all fatty acid elongase (ELO) and desaturase (DES) genes required for DHA synthesis. The reverse genetic approach and a tracing experiment using stable isotope-labeled fatty acids revealed that the ELO/DES pathway is the only DHA synthesis pathway in Parietichytrium sp. Disruption of the C20 fatty acid ELO (C20ELO) and ∆4 fatty acid DES (∆4DES) genes with expression of ω3 fatty acid DES in this thraustochytrid allowed the production of EPA and n-3docosapentaenoic acid (n-3DPA), respectively, at the highest level among known microbial sources using fed-batch culture.


2021 ◽  
Vol 18 (22) ◽  
pp. 453
Author(s):  
Azila Adnan ◽  
Giridhar Nair ◽  
Mark Lay ◽  
Janis Swan

Bacterial cellulose (BC) is an abundant polysaccharide, which is secreted by several genera of bacteria. It has remarkable characteristics, which include high purity, high tensile strength, high biocompatibility and non-toxic. The main feature that differentiates BC and plant cellulose (PC) is the absence of contaminants such as lignin, hemicellulose and pectin. However, the main drawbacks in producing BC are low yield and expensive carbon source. Due to that, this study was carried out to enhance BC volumetric productivity in fed-batch operation mode using glycerol as a carbon source. BC was produced in fill-and-draw and pulse-feed fed-batch cultures of Gluconacetobacter xylinus DSM 46604 in a 3-L bench-top bioreactor. The fed-batch fermentation trials were conducted in agitated and aerobic conditions at 30 ºC. For fill-and-draw fed-batch culture, a total of 24.2 g/L of BC accumulated in the bioreactor after 9 days, which corresponded to a yield and productivity of 0.2 g/g and 2.69 g/L/day, respectively. Pulse-feed fed-batch fermentation resulted in a yield and volumetric productivity of 0.38 g/g and 2.71 g/L/day, respectively. The pulse-feed fed-batch culture proved to be a better fermentation system for utilizing glycerol, which is a low-cost and abundant carbon source. HIGHLIGHTS Komagataeibacter species, which were formerly known as Acetobacter or Gluconacetobacter is one of the Gram-negative BC producers that secretes a large quantity of BC microfibrils extracellularly One of the main challenges in bacterial cellulose (BC) production is low productivity and high processing cost As fed-batch fermentation is one of the operation modes in bioprocess that can control the microbial growth rate, this operation mode is conducted to enhance the yield of BC, substrate consumption and also volumetric productivity Fill-and-draw and pulse feed fed-batch culture were conducted to enhance yield and volumetric productivity. The pulse-feed fed-batch culture resulted to be a favorable operation mode for utilizing glycerol, which is a low-cost and abundant carbon source GRAPHICAL ABSTRACT


PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0248382
Author(s):  
Emanuela Palomba ◽  
Valentina Tirelli ◽  
Elisabetta de Alteriis ◽  
Palma Parascandola ◽  
Carmine Landi ◽  
...  

The yeast Saccharomyces cerevisiae is a reference model system and one of the widely used microorganisms in many biotechnological processes. In industrial yeast applications, combined strategies aim to maximize biomass/product yield, with the fed-batch culture being one of the most frequently used. Flow cytometry (FCM) is widely applied in biotechnological processes and represents a key methodology to monitor cell population dynamics. We propose here an application of FCM in the analysis of yeast cell cycle along the time course of a typical S. cerevisiae fed-batch culture. We used two different dyes, SYTOX Green and SYBR Green, with the aim to better define each stage of cell cycle during S. cerevisiae fed-batch culture. The results provide novel insights in the use of FCM cell cycle analysis for the real-time monitoring of S. cerevisiae bioprocesses.


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