US 2016.0167993A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2016/0167993 A1 RAZAV-SHRAZI et al. (43) Pub. Date: Jun. 16, 2016 (54) BOCATALYST COMPOSITIONS AND 2012, provisional application No. 61/689,933, filed on PROCESSES FOR USE Jun. 15, 2012, provisional application No. 61/689,932, filed on Jun. 15, 2012, provisional application No. (71) Applicant: MICROVI BIOTECH, INC. Hayward, 61/689,930, filed on Jun. 15, 2012, provisional appli CA (US) cation No. 61/689,929, filed on Jun. 15, 2012, provi sional application No. 61/689.925, filed on Jun. 15, (72) Inventors: Fatemeh RAZAVI-SHIRAZI, 2012, provisional application No. 61/689.924, filed on Hayward, CA (US); Mohammad Ali Jun. 15, 2012, provisional application No. 61/689,923, DORRI, Milpitas, CA (US); Farhad filed on Jun. 15, 2012, provisional application No. DORRI-NOWKOORANI, Union City, 61/689,922, filed on Jun. 15, 2012, provisional appli CA (US); Ameen RAZAVI, Fremont, cation No. 61/689,921, filed on Jun. 15, 2012. CA (US) Publication Classification (21) Appl. No.: 14/935,164 (51) Int. C. (22) Filed: Nov. 6, 2015 CO2F 3/10 (2006.01) CI2N II/04 (2006.01) Related U.S. Application Data CI2N II/08 (2006.01) (52) U.S. C. (62) Division of application No. 13/918,868, filed on Jun. CPC ................ C02F3/108 (2013.01): CI2N II/08 14, 2013, now Pat. No. 9,212,358. (2013.01); C12N II/04 (2013.01); C02F (60) Provisional application No. 61/852.451, filed on Mar. 2003/001 (2013.01) 15, 2013, provisional application No. 61/851,467, filed on Mar. 8, 2013, provisional application No. (57) ABSTRACT 61/850,631, filed on Feb. 20, 2013, provisional appli The microorganism-containing biocatalysts disclosed have a cation No. 61/849,725, filed on Feb. 1, 2013, provi large population of the microorganisms irreversibly retained sional application No. 61/689,953, filed on Jun. 15, in the interior of the biocatalysts. The biocatalysts possess a 2012, provisional application No. 61/689,945, filed on Surprisingly stable population of microorganisms and have an Jun. 15, 2012, provisional application No. 61/689,943, essential absence of debris generation from metabolic activ filed on Jun. 15, 2012, provisional application No. ity of the microorganisms. The biocatalysts are composed of 61/689,940, filed on Jun. 15, 2012, provisional appli highly hydrophilic polymer and have an internal, open, cation No. 61/689,939, filed on Jun. 15, 2012, provi porous structure that promotes community phenotypic sional application No. 61/689.935, filed on Jun. 15, changes. Patent Application Publication Jun. 16, 2016 Sheet 1 of 9 US 2016/0167993 A1 FIGURE 1 Patent Application Publication Jun. 16, 2016 Sheet 2 of 9 US 2016/0167993 A1 316 3. 3. 308 3. 3. 33 FIGURE 3 s FIGURE 4 Patent Application Publication Jun. 16, 2016 Sheet 3 of 9 US 2016/0167993 A1 Si FIGURE 5 Patent Application Publication Jun. 16, 2016 Sheet 4 of 9 US 2016/0167993 A1 fia. ; 5. t isis FGURE is Patent Application Publication Jun. 16, 2016 Sheet 5 of 9 US 2016/0167993 A1 FGURE 7 Patent Application Publication Jun. 16, 2016 Sheet 6 of 9 US 2016/0167993 A1 BOREACTOR SEO ENCING EF ER3. REf “... Shi R.S.A. Šajiš. ES AEE is: R - ::::::: P, Raivial o, REMWAL PO, RELEASE EE EE S Ps SE EC ii. i. f PO, REE, AL FO, RELEASE A SEith PO REC.. O & Polishing Fix A.Y.E. s iii. is ENERA P, REFEO.A. PC. R.E.O. At REE 2, RELEASE i E LE: is: s: is: GE P. RELEASE EERA. ; Relial : as: g: is: ski EB GEE PO, RELEASE P. REritoval Pl, EwAwa. E i. ce FIGURE 8 Patent Application Publication Jun. 16, 2016 Sheet 7 of 9 US 2016/0167993 A1 3. FGRE 9 Patent Application Publication Jun. 16, 2016 Sheet 8 of 9 US 2016/0167993 A1 FIGURE 10 Patent Application Publication Jun. 16, 2016 Sheet 9 of 9 US 2016/0167993 A1 i. 156 -- FIGURE 1 2 E. - E. FGRE US 2016/0167993 A1 Jun. 16, 2016 BOCATALYST COMPOSITIONS AND improving metabolic processes for industrial use and expand PROCESSES FOR USE ing the variety of metabolic process alternatives to chemical syntheses and degradations. CROSS-REFERENCE TO RELATED 0024 Numerous types of process techniques have been APPLICATIONS proposed for anabolic and catabolic bioconversions. These 0001 Priority is claimed to U.S. Provisional Patent Appli processes include the use of suspended microorganisms, i.e., cations Nos.: planktonic processes. Also, process techniques have been 0002 61/689,921, filed on Jun. 15, 2012: disclosed where the microorganisms are located on or within 0003. 61/689,922, filed on Jun. 15, 2012: a solid Support. 0004. 61/689,923, filed on Jun. 15, 2012: 0025 Workers are faced with various challenges in 0005 61/689,924, filed on Jun. 15, 2012: improving metabolic processes and in providing metabolic 0006 61/689,925, filed on Jun. 15, 2012: processes that are sufficiently economically viable to be of 0007. 61/689,929, filed on Jun. 15, 2012: commercial interest. Some problems may be inherent with 0008. 61/689,930, filed on Jun. 15, 2012: the feedstock itself including the presence of toxins, phages, 0009 61/689,932, filed on Jun. 15, 2012: and adventitious competitive microorganisms. Other prob O010 61/689,933, filed on Jun. 15, 2012: lems may arise from the microorganism to be used for the 0011. 61/689,935, filed on Jun. 15, 2012: bioconversion Such as low metabolic conversion rate, low 0012 61/689,939, filed on Jun. 15, 2012: population growth rate, automutation, significant consump 0013 61/689,940, filed on Jun. 15, 2012: tion of Substrate to Support population growth, the need for 0014 61/689,943, filed on Jun. 15, 2012: inducers, co-metabolites, promoters and performance 0.015 61/689,945, filed on Jun. 15, 2012: enhancing additives, and the lack of a microorganism that has 0016. 61/689,953, filed on Jun. 15, 2012: the sought metabolic conversion. And yet further problems 0017 61/849,725, filed on Feb. 1, 2013; may arise from the process used for the bioconversion Such as 0018 61/850,631, filed on Feb. 20, 2013; costs in recovering bioproducts from an aqueous fermenta 0019 61/851,467, filed on Mar. 8, 2013; and tion broth. Especially with Supported microorganisms, prob 0020. 61/852.451, filed on Mar. 15, 2013, lems can arise from instability of the biofilms, including their each of which is hereby incorporated by reference in its physical degradation; overgrowth of the population of micro entirety. A right is hereby reserved to have patentability deter organisms causing Suffocation; sloughing off of the microor minations made on the basis of the applicable sections of ganisms from the Support; and susceptibility to competitive Public Law 112-29. microorganisms. Additionally, metabolic processes are char acterized as generating solid debris from dead or lysed cells, STATEMENT REGARDING FEDERALLY and the debris needs to be accommodated in the process to SPONSORED RESEARCH ORDEVELOPMENT remove these solids. In some instances, the debris have value 0021. The use of biocatalysts for the continuous degrada as feed Supplements such as distillers grains from the manu tion of 1,4-dioxane in ultralow concentrations in water was facturing of ethanol, but in other metabolic processes such as first reduced to practice using Government Support under for the treatment of municipal waste water, costs may have to Contract 1R43ES022123-01, awarded by the National Insti be incurred to dispose of the debris in an environmentally tutes of Health. The Government has certain rights thereto. acceptable manner. Genetic engineering, which has been pro posed to overcome one or more of these problems, can itself FIELD OF THE INVENTION be problematic. 0022. This invention pertains to novel biocatalysts and 0026 Microorganisms, including but not limited to, bac their use. teria, archaea, fungi and algae, are capable of becoming attached or adhered to a surface. Studies have been conducted BACKGROUND pertaining to the effect of a change from planktonic growth to 0023 Metabolic processes have long been proposed for growth of microorganisms on Surfaces, including the forma anabolic and catabolic bioconversions. Microorganisms of tion of biofilms on surfaces. A number of workers have inves various types have been proposed for these bioconversions tigated preventing or degrading biofilms in an animal or and include bacteria and archaea, both of which are prokary human body to enhance the efficacy of antibiotic treatments otes; fungi, and algae. Metabolic processes are used by to cure the animal or human body. nature, and some have been adapted to use by man for mil 0027 Tuson, et al., in “Bacteria-surface Interactions', lennia for anabolic and catabolic bioconversions ranging SoftMatter, Vol. 1, issue 608 (2013) citable as DOI: 10.1039/ from culturing yogurt and fermentation of Sugars to produce c3sm27705d, provide a review of work in the field of bacte alcohol to treatment of water to remove contaminants. Meta rial-Surface interactions. The authors describe the processes bolic processes offer the potential for low energy consump involved in attaching a microorganism to a Surface and recite tion, high efficiency bioconversions in relatively inexpensive that attachment to Surfaces causes phenotypic Switches in the processing equipment and thus may be and are often viable cells and that the surface can provide benefits to the attached alternatives to chemical synthesis and degradation methods. cells. The authors recite that organic matter can concentrate at Often anabolic processes can use raw materials that are pre horizontal Surfaces stimulating growth of bacteria associated ferred from a renewable or environmental standpoint but are with the Surface, and increasing Substrate surface area pro not desirable for chemical synthesis, e.g., the conversion of vides more area on which nutrients can absorb, enabling cells carbon dioxide to biofuels and other bioproducts.