High cell density co-culture for production of recombinant hydrolases

Silva, M.R.; Severo, M.G.; Delabona, P. S.; Ruller, Roberto; Pradella, J. G. C.; Gonçalves, Viviane Maimoni; Freitas, Sindélia

doi:10.1016/j.bej.2012.11.008

Cited by 5 publications

(7 citation statements)

References 30 publications

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“…On the other hand, the yield values for the main fermenter were estimated by applying a 20% reduction to the batch yield values; as such, they were equal to 0.40 g/g for glucose and 0.36 g/g for glycerol. Based on our experience, the biomass yield can be reduced by as much as 50% in fed-batch mode compared to batch culture mode (30). Furthermore, these lower yield values are in reasonable agreement with those obtained by Wyre and Overton (34), who employed somewhat similar conditions (a fed-batch culture of E. coli with IPTG induction).…”

Section: Culture Media and Fermentation Conditionssupporting

confidence: 89%

“…However, the value of specific growth rate derived in the present work is also higher than that obtained by Silva et al (30), who cultivated the BL21(DE3) strain with pET28aderived plasmids in a bioreactor under the same conditions and using the same medium, except for employing a higher concentration of glucose (µ = 0.56-0.59 h -1 ). More generally, the maximum specific growth rate achieved here is in line with values obtained by various strains of E. coli growing at 37 °C on defined medium with glucose as the limiting substrate, which fall in the range of 0.5-1.0 h -1 (269).…”

Section: Growth Kinetics In Hdf Medium Using a Bioreactorcontrasting

confidence: 73%

“…However, the use of plasmid DNA for rProtein production faces a significant challenge: if left unchecked, recombinant cells tend to lose their plasmids and, as a result, rProtein productivity declines precipitously. This phenomenon occurs because the very expression of rProteins, and even plasmid maintenance and replication by themselves, constitute a metabolic burden for the microorganism, so that cells that lose plasmid molecules during division tend to outcompete plasmid-harboring cells (25,30,31). To prevent this problem, plasmid stability is usually ensured by adding an antibiotic to the culture medium while having the corresponding antibiotic resistance gene inserted into the plasmid (23,27,32).…”

Section: List Of Figuresmentioning

confidence: 99%

“…17,19,30,51,63,91,[160][161][162][163]; endowing cellulolytic organisms such as T. reesei or Clostridium with lacking enzyme activities or better enzymes, through genetic and protein engineering(4,6,155,(164)(165)(166)(167)(168)(169)12,13,16,42,48,86,142,143); co-culturing T. reesei with good producers of lacking enzymes, such as Aspergillus sp (16,30,48,142,163,164,170)…”

mentioning

confidence: 99%

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Development and in silico evaluation of an expression platform based on E.coli for the production of a recombinant beta-glucosidase.

Ferreira¹,

Azzoni²

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Section: Culture Media and Fermentation Conditionssupporting

confidence: 89%

Section: Growth Kinetics In Hdf Medium Using a Bioreactorcontrasting

confidence: 73%

Section: List Of Figuresmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Development and in silico evaluation of an expression platform based on E.coli for the production of a recombinant beta-glucosidase.

Ferreira¹,

Azzoni²

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show abstract

“…The yield values for the main fermenter were estimated by applying a 20% reduction to the batch yield values. Based on our experience, the biomass yield can be reduced by as much as 50% in fed-batch mode compared to batch culture mode [ 31 ]. The glucose and glycerol values are both in reasonable agreement with those obtained by Wyre and Overton [ 32 ], who used somewhat similar conditions.…”

Section: Methodsmentioning

confidence: 99%

Techno-economic analysis of the industrial production of a low-cost enzyme using E. coli: the case of recombinant β-glucosidase

Ferreira

Azzoni

Freitas

2018

Biotechnol Biofuels

116

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BackgroundThe enzymatic conversion of lignocellulosic biomass into fermentable sugars is a promising approach for producing renewable fuels and chemicals. However, the cost and efficiency of the fungal enzyme cocktails that are normally employed in these processes remain a significant bottleneck. A potential route to increase hydrolysis yields and thereby reduce the hydrolysis costs would be to supplement the fungal enzymes with their lacking enzymatic activities, such as β-glucosidase. In this context, it is not clear from the literature whether recombinant E. coli could be a cost-effective platform for the production of some of these low-value enzymes, especially in the case of on-site production. Here, we present a conceptual design and techno-economic evaluation of the production of a low-cost industrial enzyme using recombinant E. coli.ResultsIn a simulated baseline scenario for β-glucosidase demand in a hypothetical second-generation ethanol (2G) plant in Brazil, we found that the production cost (316 US$/kg) was higher than what is commonly assumed in the literature for fungal enzymes, owing especially to the facility-dependent costs (45%) and to consumables (23%) and raw materials (25%). Sensitivity analyses of process scale, inoculation volume, and volumetric productivity indicated that optimized conditions may promote a dramatic reduction in enzyme cost and also revealed the most relevant factors affecting production costs.ConclusionsDespite the considerable technical and economic uncertainties that surround 2G ethanol and the large-scale production of low-cost recombinant enzymes, this work sheds light on some relevant questions and supports future studies in this field. In particular, we conclude that process optimization, on many fronts, may strongly reduce the costs of E. coli recombinant enzymes, in the context of tailor-made enzymatic cocktails for 2G ethanol production.Electronic supplementary materialThe online version of this article (10.1186/s13068-018-1077-0) contains supplementary material, which is available to authorized users.

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On the production cost of lignocellulose‐degrading enzymes

Ferreira

Azzoni

Freitas

2020

Biofuels Bioprod Bioref

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Lignocellulose is the most abundant renewable material on Earth and the primary component of agricultural wastes such as sugarcane bagasse and wheat straw. It consists of a composite material made of cellulose, hemicellulose, and lignin. Cellulose and hemicellulose can be broken down into monomers by a set of appropriate enzymes, and the resulting monomers may be used to produce a variety of fuels or chemicals through either biological or chemical routes. However, the high production cost of these lignocellulose‐degrading enzymes remains a major challenge for the use of lignocellulosic biomass as raw material. In this context, this article reviews techno‐economic analyses concerning the production of cellulases and other lignocellulose‐degrading enzymes published over the last two decades. The major characteristics of each enzyme production process are described, underscoring the similarities and differences across the various process designs. Moreover, the enzyme production costs derived from these process designs and their composition in terms of raw materials, capital‐related factors, utilities, labor costs, etc., are compared. First, this analysis reveals that most techno‐economic evaluations in the literature address either cellulase production by submerged culture with Trichoderma reesei or enzyme production by solid‐state culture with filamentous fungi. Second, this analysis shows wide cost variations across process designs but it indicates that raw materials and capital‐related costs are generally the main drivers of the enzyme production cost. Furthermore, this assessment corroborates the importance of process parameters, such as product yield, production titer, and volumetric productivity, in the process economics of enzyme production. © 2020 Society of Chemical Industry and John Wiley & Sons, Ltd

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High cell density co-culture for production of recombinant hydrolases

Cited by 5 publications

References 30 publications

Development and in silico evaluation of an expression platform based on E.coli for the production of a recombinant beta-glucosidase.

Development and in silico evaluation of an expression platform based on E.coli for the production of a recombinant beta-glucosidase.

Techno-economic analysis of the industrial production of a low-cost enzyme using E. coli: the case of recombinant β-glucosidase

On the production cost of lignocellulose‐degrading enzymes

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