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

Abstract

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.

Recombinant Proteins for Industrial versus Pharmaceutical Purposes: A Review of Process and Pricing

“…This leads to a lower contaminant load in the product stream. Proteins made in E. coli frequently need to be refolded from aggregated protein (inclusion bodies) after lysis of the cells [14]. For reasons outlined above, for IRPs, non-E. coli or fungal systems are now preferred, and some of them have the capacity to secrete protein as well.…”

“…Calculations were derived from the United States Centers for Medicare and Medicaid Services (CMS) Average Sales Price (ASP) and the pharmaceutical package insert [14]. rFVIII: 7.5 × 10 3 IU/mg × 10 6 mg/kg = 7.5 × 10 9 IU/kg × $1.28/IU = $9.6 × 10 9 /kg rFVII: $2.07/mcg × 10 9 mcg/kg = $2.07 × 10 9 /kg Rituximab: $95/mL × 1 mL/10 mg × 10 6 mg/kg = $9.5 × 10 6 /kg rHGH: $55/0.4 mg × 10 6 mg/kg = $1.37 × 10 8 /kg eculizumab: $230/mL × 1 mL/10 mg × 10 6 mg/kg = $2.3 × 10 7 /kg rHepatitis B surface antigen: $27/5 mcg × 10 9 mcg/kg = $ 5.4 × 10 9 /kg The price of cellulase and glucosidase were determined from the cited references for Table 1 [13,14].…”

“…Despite similarities in manufacturing techniques used to produce PRPs and IRPs, drastic differences in retail pricing exist. IRPs often sell for tens of dollars per kilogram (kg) while PRPs can sell for billions of dollars per kg (Table 1) [13][14][15].…”

Aspergillus: A Powerful Protein Production Platform

“…For the production of non-medicinal proteins, a more economical approach is usually followed, using either bacterial or fungal production hosts [1,6,7]. While bacteria are often suitable for smaller proteins that do not require complex post-translational modifications, production of larger and more complex proteins is usually performed in yeast, e.g., Pichia pastoris [8].…”

Imobilização de β-glicosidase de soja em bucha vegetal (Luffa cylindrica) utilizando trimetafosfato de sódio como ativador

“…2 Também são utilizadas pela indústria de combustíveis em matérias-primas renováveis para conversão de biomassa lignocelulósica em açúcares fermentecíveis. 3 Porém, seu uso ainda é limitado em razão da instabilidade de sua estrutura, dificuldade de separação da enzima do produto final e a impossibilidade de sua recuperação para aplicações repetidas. 4,5 Uma alternativa para esse problema seria a sua imobilização.…”