Manufacturing demonstration of microbially mediated zinc sulfide nanoparticles in pilot-plant scale reactors


The thermophilic anaerobic metal-reducing bacterium Thermoanaerobacter sp. X513 efficiently produces zinc sulfide (ZnS) nanoparticles (NPs) in laboratory-scale (≤ 24-L) reactors. To determine whether this process can be up-scaled and adapted for pilot-plant production while maintaining NP yield and quality, a series of pilot-plant scale experiments were performed using 100-L and 900-L reactors. Pasteurization and N2-sparging replaced autoclaving and boiling for deoxygenating media in the transition from small-scale to pilot plant reactors. Consecutive 100-L batches using new or recycled media produced ZnS NPs with highly reproducible ~2-nm average crystallite size (ACS) and yields of ~0.5 g L−1, similar to the small-scale batches. The 900-L pilot plant reactor produced ~320 g ZnS without process optimization or replacement of used medium; this quantity would be sufficient to form a ZnS thin film with ~120 nm thickness over 0.5 m width × 13 km length. At all scales, the bacteria produced significant amounts of acetic, lactic, and formic acids, which could be neutralized by the controlled addition of sodium hydroxide without the use of an organic pH buffer, eliminating 98 % of the buffer chemical costs. The final NP products were characterized using XRD, ICP-OES, TEM, FTIR, PL, DLS, HPLC, and C/N analyses, which confirmed that the growth medium without organic buffer enhanced the ZnS NP properties by reducing carbon and nitrogen surface coatings and supporting better dispersivity with similar ACS.

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The authors gratefully acknowledge the support of the US Department of Energy (DOE), Advanced Manufacturing Office, Low Temperature Material Synthesis Program (CPS 24762). FTIR analysis performed by M.K. Kidder was supported by the US Department of Energy, Office of Science, Basic Energy Sciences, Chemical Sciences, Geosciences, and Biosciences Division. ORNL is managed by UT-Battelle, LLC, for DOE under contract DE-AC05-00OR22725. The authors thank Dr. C.B. Jacobs at ORNL for constructive discussion, Dr. J, Zhu for TEM images and S.R. Cline and J.P. Dugger for assistance designing the pilot plant and safety procedures. We also thank Stout Tanks & Kettles, LLC, for the customized reactor design and construction.

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Correspondence to Ji-Won Moon.

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This manuscript has been authored by UT-Battelle, LLC under Contract No. DE-AC05-00OR22725 with the US Department of Energy. The United States Government retains and the publisher, by accepting the article for publication, acknowledges that the United States Government retains a non-exclusive, paid-up, irrevocable, worldwide license to publish or reproduce the published form of this manuscript, or allow others to do so, for United States Government purposes. The Department of Energy will provide public access to these results of federally sponsored research in accordance with the DOE Public Access Plan (http://

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Moon, JW., Phelps, T.J., Fitzgerald Jr, C.L. et al. Manufacturing demonstration of microbially mediated zinc sulfide nanoparticles in pilot-plant scale reactors. Appl Microbiol Biotechnol 100, 7921–7931 (2016).

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  • Pilot plant reactor
  • Microbially mediated manufacturing
  • Zinc sulfide nanoparticles
  • Scalability