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WO 2014/105944 Al 3 July 2014 (03.07.2014) P O P C T

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WO 2014/105944 Al 3 July 2014 (03.07.2014) P O P C T (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2014/105944 Al 3 July 2014 (03.07.2014) P O P C T (51) International Patent Classification: (74) Agent: SHARP, Jeffrey, A.; 2040 Avenue C, Bethlehem, C21B 11/00 (2006.01) C22B 11/10 (2006.01) Pennsylvania 18017 (US). (21) International Application Number: (81) Designated States (unless otherwise indicated, for every PCT/US20 13/077790 kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, (22) Date: International Filing BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, 26 December 2013 (26. 12.2013) DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, (25) Filing Language: English HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, ME, (26) Publication Language: English MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, (30) Priority Data: OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, 61/746,7 10 28 December 201 2 (28. 12.20 12) US SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, (71) Applicant: FLSMIDTH A S [DK/DK]; 77 Vigerslev Alle, ZW. DK-2500 Valby (DK). (84) Designated States (unless otherwise indicated, for every (72) Inventor; and kind of regional protection available): ARIPO (BW, GH, (71) Applicant (for US only): ROCKS, Sara S. [US/US]; GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, SZ, TZ, 2665 E. Robidoux Rd, Sandy, Utah 84093 (US). UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, (72) Inventor: CHAIKO, David J.; 10952 Langford Ln Apt EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, 2102, South Jordan, Utah 84095 (US). MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, [Continued on nextpage] (54) Title: USE OF ENZYMES FOR RECOVERING A METAL FROM A METAL-CONTAINING ORE (57) Abstract: Enzyme-based leaching agents for leaching metals from metal- 100 containing ores. Methods for recovering metals from a metal-containing ores us ing such enzyme-based leaching agents are also described. Enzymes that are act -110 ive under leaching conditions (e.g., low pH, high ionic strength, high temperat Mine Ore ures) may be isolated from microorganisms. In addition, the activity of enzymes that are identified as being active under leaching conditions may be improved through protein engineering techniques such as, but not limited to, random -120 Comminution mutagenesis, site directed mutagenesis, directed evolution, combinatorial tech niques, and the like. 130 Floatation Leaching -140 Remove -150 Impurities Extraction -wo Electrowinning -170 © © o w o 2014/105944 Illlll II Hill lllll Hill llll III III Hill Hill lllll lllll lllll ilimn i i llll TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, Published: KM, ML, MR, NE, SN, TD, TG). with international search report (Art. 21(3)) Declarations under Rule 4.17: before the expiration of the time limit for amending the — as to applicant's entitlement to apply for and be granted claims and to be republished in the event of receipt of a patent (Rule 4.1 7(H)) amendments (Rule 48.2(h)) — of inventorship (Rule 4.17(iv)) USE OF ENZYMES FOR RECOVERING A METAL FROM A METAL- CONTAINING ORE CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is an international PCT application which claims priority to United States Provisional Patent Application Serial No. 61/746,710 filed on 28 December 2012. BACKGROUND [0002] Many of the world's metal containing ores contain only small amounts (e.g., by percent weight) of desirable metals. In order to make such ores commercially viable, it is often necessary to leach (i.e., extract) the metal from the ore and concentrate it prior to further processing. [0003] For example, over 90% the world's mine copper is currently obtained from copper sulfide ore processing. The most important copper sulfide species present in ores are chalcopyrite, bornite chalcosite, covellite, tenantite and enargite, of which chalcopyrite is the species found in most relative abundance and, therefore, the one of greatest economic interest. High-grade copper ores may only contain about 2% copper by weight and low-grade ores, which may be commercially viable nonetheless, may contain less than 1% copper by weight. [0004] Copper sulfide ore processing is sustained by technologies based on physical and chemical processes associated with mineral crushing, grinding and flotation, followed by fusion-conversion of concentrates and electrolytic refining of metal. In practice, over 70% of copper is produced through the described route - known as the conventional route - which is limited to high and medium grade ores, according to the specific characteristics of deposits and of ore processing plants. [0005] On the other hand, ores in which copper is present in the form of oxide species (easily soluble in acid) are processed by means of acid leaching processes, followed by solvent extraction processes and electro-winning of the metal, in what is known as copper winning through hydrometallurgy. This route is very attractive due to its lower operation and investment costs when compared to conventional technologies, as well as to its lower environmental impact. Nevertheless, applications of this technology are limited to oxide ores, or to copper sulfide mixed ores in which metal is present in the form of secondary sulfides (chalcosite and covellite) that are acid soluble in the presence of an energetic oxidizing agent catalyzed by microorganisms. [0006] As an alternative to the processes described above, leaching of minerals may be accomplished in the presence of micro-organisms that enhance the leaching kinetics. However, the leaching environments are difficult for microorganisms due to the low pH, high ionic strength, and high temperatures. In fact, all hydrometallurgical processing conditions can be incredibly harsh to microorganisms. In some instances, extremophiles (i.e., bacteria that thrive under extreme conditions) may be used in bioleaching. Nevertheless, bioleaching is inherently inefficient because, for example, much of the organisms' energy must be expended by the organisms in life processes unrelated to mineral recovery, the organisms must be supplied with nutrients, many of which are incompatible with mineral processing and recovery, and leaching may tend to kill microorganisms or suppress their growth due to harsh environments (e.g., low pH, high ionic strength, high temperatures, etc.). SUMMARY [0007] Described herein are enzyme-based leaching agents that can be used to leach metals from metal-containing ores. Methods for recovering metals from metal- containing ores using such enzyme-based leaching agents are also described. Enzymes that are active under leaching conditions (e.g., low H, high ionic strength, high temperatures) may be isolated from microorganisms that are able to thrive under such conditions. In addition, the activity of enzymes that are identified as being active under leaching conditions may be improved through protein engineering techniques such as, but not limited to, random mutagenesis, site directed mutagenesis, directed evolution, combinatorial techniques, and the like. [0008] In an embodiment, a leaching agent for recovering a metal from a metal- containing ore is described. The leaching agent includes an acid and at least one enzyme associated with the acid and capable of promoting (e.g., enhancing) leaching metal from the metal-containing ore. For example, the leaching agent may be substantially abiotic. In one embodiment, the at least one enzyme may include a native enzyme or a recombinant enzyme that is isolated from or derived from a microorganism. [0009] In another embodiment, a method of recovering a metal from a metal- containing ore is described. The method includes (1) contacting the ore with a leaching agent that includes at least one enzyme, wherein the leaching agent is substantially abiotic, (2) performing a leaching process with the leaching agent to leach the metal from the metal-containing ore, (3) producing at least one of a solid or liquid leachate from the leaching process, wherein the leachate includes the metal from the metal- containing ore, and (4) recovering the metal from the leachate. [0010] In a more specific embodiment, the method may include a method of recovering copper from a copper sulfide-containing ore. Such a method includes (1) contacting the copper sulfide-containing ore with an acidic, substantially abiotic leaching agent that includes Fe(III) and at least one enzyme capable of oxidizing Fe(II) to Fe(III), (2) performing a leaching process to leach the copper from the copper sulfide-containing ore, (3) producing at least one of a solid or liquid leachate from the leaching process, wherein the leachate includes the copper from the copper sulfide- containing ore, and (4) recovering copper metal from the leachate. [0011] In one embodiment, the at least one enzyme may include at least one of rusticyanin or cytochrome C442. In one embodiment, the copper sulfide-containing ore may include at least one of copper sulfide (chalcocite and covellite) or copper iron sulfide (chalcopyrite and bornite). Other metals that can be recovered using the methods described herein include, but are not limited to, molybdenum, gold, silver, and nickel. [0012] In an embodiment, a screening assay may be performed to identify the at least one enzyme that is suitable for leaching the metal. In an embodiment, crystalline nano-scale mineral particles may be pre-synthysized that represents, or otherwise approximate a composition of the metal-containing ore.
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