Transforming biochemical engineering with cell‐free biology

Swartz, James R.

doi:10.1002/aic.13701

Cited by 107 publications

(98 citation statements)

References 35 publications

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“…DNA-dependent in vitro protein synthesis is becoming a popular technology for cell-free biology (Swartz, 2012). Linear or circular DNA programs are now executed in cell-free transcription-translation (TX-TL) mixtures for biomedical applications (He and Taussig, 2001), nanotechnology (Daube and Bar-Ziv, 2013), synthetic biology (Chappell et al, 2013;Iyer and Doktycz, 2014;Lentini et al, 2013;Niederholtmeyer et al, 2013;Shin and Noireaux, 2012;Sokolova et al, 2013;Sun et al, 2013) and metabolic engineering (Bujara et al, 2010;Ye et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

A cost-effective polyphosphate-based metabolism fuels an all E. coli cell-free expression system

Caschera¹,

Noireaux²

2015

Metabolic Engineering

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Section: Introductionmentioning

confidence: 99%

A cost-effective polyphosphate-based metabolism fuels an all E. coli cell-free expression system

Caschera¹,

Noireaux²

2015

Metabolic Engineering

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“…Two ways for constructing cell-free biosystems are whole crude cell extracts and purified enzymes. The former is easily prepared but most enzyme components may not last long and the existing undesired pathways may cause side reactions [4,[104][105][106]. The latter requires more labor and cost for enzyme preparation but it has better access control and easy optimization of numerous building blocks with different characteristics.…”

Section: Strategies In Cell-free Biosystems For Biomanufacturingmentioning

confidence: 98%

“…Crude cell extracts have been used for cell-free protein synthesis for a long time due to its easy preparation and low costs [4,104,143,147]. However, the other remaining cellular enzymes may cause some side reactions.…”

Section: Treated Cell Extractsmentioning

confidence: 99%

“…Further tuning of redox enzymes is needed due to its important application. Although the importance of redox enzyme engineering is recognized more and more to have a place in the future of biomanufacturing [9,104,145,146], redox enzyme engineering remains in its early stages because there is no framework or a general rule for engineering redox enzymes with nonnatural cofactors [59].…”

Section: Redox Enzyme Engineeringmentioning

confidence: 99%

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Cell-free Biosystems in the Production of Electricity and Bioenergy

Zhu

Tam²,

Zhang

2013

Fundamentals and Application of New Bioproduction Systems

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: Increasing needs of green energy and concerns of climate change are motivating intensive R&D efforts toward the low-cost production of electricity and bioenergy, such as hydrogen, alcohols, and jet fuel, from renewable sugars. Cell-free biosystems for biomanufacturing (CFB2) have been suggested as an emerging platform to replace mainstream microbial fermentation for the cost-effective production of some biocommodities. As compared to whole-cell factories, cell-free biosystems comprised of synthetic enzymatic pathways have numerous advantages, such as high product yield, fast reaction rate, broad reaction condition, easy process control and regulation, tolerance of toxic compound/product, and an unmatched capability of performing unnatural reactions. However, issues pertaining to high costs and low stabilities of enzymes and cofactors as well as compromised optimal conditions for different source enzymes need to be solved before cell-free biosystems are scaled up for biomanufacturing. Here, we review the current status of cell-free technology, update recent advances, and focus on its applications in the production of electricity and bioenergy.

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“…In vitro synthetic biology enables the rapid construction of nonnatural enzymatic pathways and often has more appealing advantages, such as higher product yields, faster reaction rates, and better tolerances to toxic compounds, than those mediated by living organisms (10,(23)(24)(25)(26)(27)(28)(29)(30)(31). Here we design a cell-free biosystem composed of a synthetic enzymatic pathway that can transform solid cellulose into amylose in high yields.…”

mentioning

confidence: 99%

Enzymatic transformation of nonfood biomass to starch

You¹,

Chen

Myung³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

148

123

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The global demand for food could double in another 40 y owing to growth in the population and food consumption per capita. To meet the world's future food and sustainability needs for biofuels and renewable materials, the production of starch-rich cereals and cellulose-rich bioenergy plants must grow substantially while minimizing agriculture's environmental footprint and conserving biodiversity. Here we demonstrate one-pot enzymatic conversion of pretreated biomass to starch through a nonnatural synthetic enzymatic pathway composed of endoglucanase, cellobiohydrolyase, cellobiose phosphorylase, and alpha-glucan phosphorylase originating from bacterial, fungal, and plant sources. A special polypeptide cap in potato alpha-glucan phosphorylase was essential to push a partially hydrolyzed intermediate of cellulose forward to the synthesis of amylose. Up to 30% of the anhydroglucose units in cellulose were converted to starch; the remaining cellulose was hydrolyzed to glucose suitable for ethanol production by yeast in the same bioreactor. Next-generation biorefineries based on simultaneous enzymatic biotransformation and microbial fermentation could address the food, biofuels, and environment trilemma.bioeconomy | food and feed | synthetic amylose | in vitro synthetic biology | cell-free biomanufacturing T he continuing growth of the population and food consumption per capita means that the global demand for food could increase by 50-100% by 2050 (1, 2), and ∼30% of the world's agricultural land and 70% of the world's fresh water withdrawals are being used for the production of food and feed to support 7 billion people (3, 4). Starch is the most important dietary component because it accounts for more than half of the consumed carbohydrates, which provide 50-60% of the calories needed by humans. Starch is composed of polysaccharides consisting of a large number of glucose units joined together primarily by alpha-1,4-glycosidic bonds and alpha-1,6-glycosidic bonds. Linear-chain amylose is more valuable than branched amylopectin because it can be used as a precursor for making high-quality transparent, flexible, low-oxygen-diffusion plastic sheets and films (5, 6); tailored functional food or additives for lowering the risk of serious noninfectious diseases (e.g., diabetes and obesity) (7, 8); and a potential high-density hydrogen carrier (9-11). Also, it is easy to convert linear amylose to branched amylopectin by using alphaglucan-branching glycosyltransferase (12).Cellulose, a linear glucan linked by beta-1,4-glycosidic bonds, is the supporting material of plant cell walls and the most abundant carbohydrate on Earth. The annual resource of cellulosic materials is ∼40 times greater than the starch produced by crops cultivated for food and feed. In addition, (perennial) cellulosic plants and dedicated bioenergy crops can grow on low-quality land, even on marginal land, and require fewer inputs such as fertilizers, herbicides, pesticides, and water, whereas annual high-productivity starch-rich crops require high-qual...

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Transforming biochemical engineering with cell‐free biology

Cited by 107 publications

References 35 publications

A cost-effective polyphosphate-based metabolism fuels an all E. coli cell-free expression system

A cost-effective polyphosphate-based metabolism fuels an all E. coli cell-free expression system

Cell-free Biosystems in the Production of Electricity and Bioenergy

Enzymatic transformation of nonfood biomass to starch

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