Abstract:
A wastewater treatment process elicits microorganisms to convert a waste stream/organic resource to intracellular biopolymer polyhydroxyalkanoate (PHA). The process includes (i) waste stream/organic resource composition feed criteria, (ii) configuration coupled with operational parameters, and (iii) PHA-laden biomass separation and stabilization. A waste stream/organic resource capable of producing enhanced levels of PHA may be selected based on a combination of criteria, which may include short chain fatty acid concentration, protein concentration, polysaccharides concentration, and total suspended solids concentration. The waste stream is introduced into an aeration basin upon a specific configuration and operated under various parameter combinations for selecting/enriching microorganisms capable of producing PHA. The PHA-laden biomass is separated and stabilized for downstream PHA related product beneficial uses. The present process achieves concurrent wastewater treatment and PHA production, where PHA level (of more than 10% on a cell-weight basis) otherwise could not be obtained.

Description:
BACKGROUND 
       [0001]    Polyhydroxyalkanoates (PHAs) are biologically derived polymers (or bioplastic) synthesized as intracellular storage materials by microorganisms metabolizing renewable organic carbon sources. The physical properties of PHA polymers are similar to those of conventional plastics (such as polypropylene (PP) and polyethylene (PE)). In contrast with traditional petroleum-based plastics, biomass-derived PHAs are generated from renewable carbon resources and are 100% biodegradable following disposal. Experts within the field consider PHAs as a potential “green” substitute to conventional plastics. 
       SUMMARY 
       [0002]    Embodiments of the invention are defined by the claims below, not this summary. A high-level overview of various aspects of the invention are provided here for that reason, to provide an overview of the disclosure, and to introduce a selection of concepts that are further described in the detailed description section below. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used as an aid in isolation to determine the scope of the claimed subject matter. 
         [0003]    Embodiments of the present invention relate to enhanced polyhydroxyalkanoates (PHAs) production during wastewater treatment. A waste stream/organic resource that is suitable for increased PHA production may initially be identified based on a combination of constituents criteria. The criteria may include total suspended solids, short chain fatty acids concentration, protein concentration, and polysaccharides concentration. In some embodiments, the waste stream/organic resource may optionally be pre-treated to provide the constituents criteria. The waste stream/organic resource is introduced into an aeration basin that may be operated under conditions for selecting and enhancing microorganisms capable of accumulating PHA. PHA-laden biomass is separated and stabilized for beneficial PHA use. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0004]    Embodiments of the present invention are described in detail below with reference to the attached drawing figures, and wherein: 
           [0005]      FIG. 1  illustrates a schematic diagram of PHA production during waste stream reclamation/organic resource recovery in accordance with an embodiment of the present invention; and 
           [0006]      FIG. 2  illustrates a schematic diagram of PHA production during waste stream reclamation/organic resource recovery using a return stream reactor in accordance with another embodiment of the present invention. 
       
    
    
     DETAILED DESCRIPTION 
       [0007]    The subject matter of the present invention is described with specificity herein to meet statutory requirements. However, the description itself is not intended to limit the scope of this patent. Rather, the inventors have contemplated that the claimed subject matter might also be embodied in other ways, to include different steps or combinations of steps similar to the ones described in this document, in conjunction with other present or future technologies. Moreover, although the terms “step” and/or “block” may be used herein to connote different elements of methods employed, the terms should not be interpreted as implying any particular order among or between various steps herein disclosed unless and except when the order of individual steps is explicitly described. 
         [0008]    Embodiments of the present invention are generally directed to producing increased polyhydroxyalkanoates (PHAs) during treatment of wastewater by converting the organic matter in the wastewater into intracellular PHA during an activated sludge (AS) process. Accordingly, the process achieves simultaneous wastewater treatment performance and increased PHA production within a bioreactor. In conventional/regular wastewater treatment process, the PHA in AS is typically less than 2.5% on a cell-weight basis. In contrast, embodiments of the present invention may produce AS with PHA above 10% on a cell-weight basis. 
         [0009]    While conventional AS processes are designed based on biological growth physiology in which environmental conditions are provided for microorganisms to convert organics in wastewater to biomass growth, embodiments of the present invention provide a process that is based on a storage-oriented perspective. Generally, environmental conditions govern the fate and conversion of organic matter by microbial consortia as either storage (i.e., PHA or glycogen accumulation) or growth response (i.e., biomass assimilation). Embodiments of the present invention focus on recovering the organic carbon by storage as the intracellular biopolymer, PHA. This is provided by selecting a waste stream with particular constituent characteristics and employing operational criteria that foster PHA storage-oriented AS (instead of growth-oriented AS as in traditional wastewater treatment). Subsequently, the excess AS that is commonly wasted in a traditional WWTP can be collected and treated for the downstream beneficial use as the AS contains a substantially higher amount of PHA. 
         [0010]    In accordance with embodiments of the present invention, a waste stream that is suitable for enhanced PHA production is initially selected. In particular, this may include evaluating the constituents of existing waste streams to identify a rapidly biodegradable, high carbon-content waste stream having PHA precursor metabolites such that the waste stream promotes a rapid mass transport of the substrate within microbial cells, which subsequently triggers the microbial storage-response metabolism. The constituents of a waste steam evaluated may include the short chain fatty acids (SCFAs) concentration, the polysaccharides concentration, and the protein concentration. 
         [0011]    SCFAs are primary precursor metabolites for PHA production. Accordingly, the waste stream should have a minimum level of SCFAs to promote PHA production. In some embodiments, a waste stream with a SCFAs concentration greater than 1 mM is preferred. 
         [0012]    Polysaccharides and protein each provide competition with (and therefore interfere with) the production of PHA. In particular, given that polysaccharides and protein are two main cell components, a relatively high polysaccharide and protein content favors assimilation into microbial biomass and, therefore, fewer PHA storage polymers. The environmental conditions provided in the main reactor enrich microbial storage capacity. However, high polysaccharide content waste streams may result in the storage of glycogen (i.e., a type of polysaccharides) instead of PHAs. In some embodiments of the present invention, a waste stream having a polysaccharides concentration less than 6 mM and a protein concentration less than 1 mM is preferred. 
         [0013]    The waste stream may further be evaluated for compounds toxic to bacteria. Preferably, the waste stream includes little to no toxic compounds to provide a non-toxic environment for microbes to survive and generate PHA. 
         [0014]    In some embodiments, a waste stream may be selected with constituents that do not have a satisfactory SCFAs concentration, polysaccharides concentration, and/or protein concentration. In such embodiments, a pre-treatment process, such as fermentation of solids, may be employed to provide a pre-treated waste stream that meets the constituent criteria discussed above. 
         [0015]    Solids may also interfere with reactor operation and “dilute” PHA content in the end products (i.e., PHA-laden biomass) while harvesting. Accordingly, in some embodiments, a waste stream is selected with minimum solids interference. Preferably, the waste stream has a total suspended solids (TSS) concentration less than 200 mg/L. If a waste stream is selected that has an undesirable level of solids (e.g., a TSS concentration more than 200 mg/L), a pre-treatment process, such as clarification or filtration, may be employed to produce a pre-treated waste stream with reduced solids to minimize solids interference with PHA production. 
         [0016]    With reference now to  FIG. 1 , a schematic view is provided that illustrates a wastewater treatment process  100  that provides concurrent PHA production in accordance with an embodiment of the present invention. As shown in  FIG. 1 , an influent waste stream  102  is treated in a manner to encourage PHA production during the treatment process. In some embodiments, the influent waste stream  102  is an untreated waste stream that includes the constituent characteristics (e.g., satisfactory SCFAs concentration, polysaccharides concentration, protein concentration, and suspended solids concentration) as described hereinabove. However, in other embodiments, one or more pre-treatment processes  104  may optionally be provided to produce the influent waste stream  102 . The pre-treatment processes may include fermentation or other process to provide a satisfactory SCFAs concentration, polysaccharides concentration, and/or protein concentration. Additionally or alternatively, the pre-treatment processes  104  may include a solids removal process, such as clarification or filtration, to reduce the suspended solids concentration of the influent waste stream  102 . 
         [0017]    The influent waste stream  102  is introduced into an aeration basin  106  and mixed with return activated sludge (RAS)  108 . In some embodiments, the aeration basin  106  may be operated as a plug-flow reactor. Additionally, the aeration basin  106  is configured with operational parameters that provide sufficient reaction time and a food to microorganism (F/M) ratio for microorganisms to uptake and deplete the substrate. The configuration ensures a famine-feast regime to select and enrich microbes capable of producing PHA. Under a feast-famine dynamic feeding pattern, AS is subjected to successive periods of external substrate availability (i.e., feast stage) and unavailability (i.e., famine stage), which generates a selective pressure that ensures microorganisms capable of generating internal storage reserves have a strong competitive advantage over those without the storage capacity. In particular, during the feast stage, microorganisms uptake available external substrate and convert it into intracellular PHA. Subsequently, PHA accumulating microorganisms consume PHA as an internal carbon source for survival during famine stage (i.e., external substrate unavailability). With respect to substrate availability, a conventional growth-oriented wastewater treatment process provides a feast stage until the external substrate is depleted; after which, treated effluent is ready to discharge. As such, in accordance with embodiments of the present invention, the location at which the influent waste stream  102  is introduced (as described in further detail below) coupled with the operational parameters of the aeration basin  106  provide a famine-feast regime (i.e., an additional famine region in front of the feast region). 
         [0018]    The operational parameters used to provide the famine-feast regime to promote PHA production in some particular embodiments may include influent COD loading rate, hydraulic retention time (HRT), and F/M ratio. Generally, the influent COD loading rate may be higher than a conventional AS process. In some embodiments, the influent COD loading rate is between 800 mg/L/day and 3,500 mg/L/day. The F/M ratio may be higher than a conventional AS process; preferably, the F/M ratio is between 0.8 and 3. 
         [0019]    The HRT may be higher than a conventional AS process. In some embodiments, longer HRTs, such as 2 or more days, may be employed to further promote the famine-feast regime. However, in other embodiments, the HRT may be as low as 1.5 days. To achieve a lower HRT (i.e., 1.5 days), the process reactor can initially be operated at a higher HRT (e.g., 2 or more days) to provide an environment that optimizes selecting/enriching microorganism capable of producing PHA. Those microorganisms may then be used to seed a process reactor with lower HRTs (e.g., 1.5 days). 
         [0020]    As shown in  FIG. 1 , the RAS  108  is introduced substantially at the beginning of the aeration basin  106 , while the influent waste stream  102  is introduced further down the aeration basin  106 . In some embodiments, the influent waste stream  102  is introduced at a location that ranges from approximately ¼ to ¾ along the length of the aeration basin  106 . Introducing the RAS  108  at the beginning of the aeration basin  106  while introducing the influent waste stream  102  further down the aeration basin  106  in this manner promotes the famine-feast regime that provides an environment that favors microorganisms capable of accumulating PHA. Therefore, an additional famine region in front of feast region promotes the enrichment/selection of PHA accumulating microorganisms. Additionally, maximum PHA production occurs at the end of the aeration basin  106 /wastewater treatment (i.e. end of feast stage) as it is ready to be collected for downstream beneficial use. Meanwhile, the treated effluent is ready to discharge (since the external substrate is depleted at the end of the feast stage). 
         [0021]    A treated waste stream  110  exiting the aeration basin  106  is processed in a solids separation basin  112 , such as a clarifier, to separate AS  114  from an effluent  116 . A first portion of the AS  108  is returned to the aeration basin  106  as the RAS  108 . A second portion of the AS  118  is provided as a PHA-laden biomass to a separation and stabilization process  120 . The separation and stabilization process  120  prevents microbes from consuming PHA as an internal carbon source following harvest and thereby produces a stabilized PHA-laden biomass  122 . In some embodiments, the separation and stabilization process  120  may include dewatering (e.g., centrifugation) followed by microbial inactivation (e.g., disinfection) and an ensuing drying process. 
         [0022]    Although only a single aeration basin  106 , solids separation basin  112 , and separation and stabilization process  120  are shown in  FIG. 1 , it should be understood that multiple aeration basins, solid separation basins, and separation and stabilization processes may be operated in parallel in accordance with embodiments of the present invention. Additionally, further basins and processes (such as a SBR) not shown in  FIG. 1  may be employed. Any and all such variations are contemplated to be within the scope of embodiments of the present invention. 
         [0023]    While  FIG. 1  illustrates an embodiment employing a single reactor (i.e., aeration basin  106 ), further embodiments may employ a return stream reactor in addition to a main stream reactor to provide the famine stage and feast stage separately that promotes PHA production. Turning now to  FIG. 2 , a schematic diagram is provided showing an embodiment of a process  200  employing a return stream reactor  226  in addition to an existing aeration basin  206 . In some embodiments, an existing wastewater treatment process may be identified with an existing aeration basin (e.g., aeration basin  206 ), and the wastewater treatment process may be modified by adding a return stream reactor (e.g., return stream reactor  226 ) to provide a process (i.e., famine stage) that optimizes PHA production. 
         [0024]    In accordance with the embodiment shown in  FIG. 2 , an influent waste stream  202  is provided that has constituent characteristics as described hereinabove. In some embodiments, this may optionally include a pre-treatment process  204 , such as fermentation of solids and/or solids separation. The influent waste stream  202  and a RAS  208  are introduced into an aeration basin  206 . In some embodiments, the operational parameters of the aeration basin  206  may include an HRT may be slightly higher than or equal to a conventional AS process, whereas the F/M ratio may be higher than or equal to a conventional AS process. In some particular embodiments, the operational parameters may include an HRT of approximately 8 hours to 2 days and an F/M ratio of 0.5 to 2. 
         [0025]    A treated waste stream  210  from the aeration basin  206  is processed in a solids separation basin  212 , such as a clarifier, to separate AS  214  from an effluent waste stream  216 . A first portion of the AS  224  is introduced into a return stream reactor  226 . In some embodiments, the return stream reactor  226  may be operated under an HRT of approximately 1 to 3 days and an F/M ratio of approximately 0.1 to 0.4. 
         [0026]    A second portion of the activate sludge  218  is provided as a PHA-laden biomass to a separation and stabilization process  220 . Similar to that discussed above with reference to  FIG. 1 , the separation and stabilization process  220  prevents microbes from consuming PHA as an internal carbon source following harvest and thereby produces a stabilized PHA-laden biomass  222 . In some embodiments, the separation and stabilization process  220  may include dewatering (e.g., centrifugation) following by microbial stabilization (e.g., disinfection) and an ensuing drying process. 
       EXAMPLE 
       [0027]    Embodiments of the present invention will now be further illustrated by the following, non-limiting examples. 
       Example 1 
       [0028]    An industrial waste stream (e.g., high-strength COD) was found to be with COD of 2,500 mg/L, SCFAs of 1.2 mM, polysaccharides of 0.8 mM, protein of 0.4 mM, and TSS of 500 mg/L. As this waste stream met the constituents criteria with the exception of TSS, solid separation was employed as a pre-treatment process to reduce solids to a desirable level of TSS less than 200 mg/L in a pre-treated waste stream. The pre-treated waste stream was then introduced at a location approximately one-third along the length of an aeration basin. The aeration basin was operated under a HRT of 3 days and F/M ratio of 1.5. RAS was introduced at the beginning of the aeration basin. In this manner, the aeration basin provided a famine-feast regime that enriched/selected microorganisms capable of accumulating PHA. A treated waste stream exiting the aeration basin was processed in a clarifier to separate AS from an effluent. A first portion of the AS was returned to the aeration basin (as noted above). A second portion of the AS was treated using centrifugation followed by disinfection and an ensuing drying process to provide a stabilized PHA-laden biomass. Meanwhile, the effluent was ready to discharge. 
       Example 2 
       [0029]    As a prophetic example of an embodiment employing a return stream reactor in addition to an aeration basin, a municipal wastewater treatment plant wastewater (e.g., low-strength COD waste stream) with COD of 250 mg/L, SCFAs of 0.2 mM, polysaccharides of 0.3 mM, protein of 0.2 mM, and TSS of 80 mg/L after existing primary sedimentation is processed. As this waste stream is not in a preferred range of COD and SCFAs concentration, a fermentation pre-treatment process is employed as a pre-treatment (e.g., primary solids fermentation) to increase COD and SCFAs to a desirable level of COD loading more than 800 mg/L/d and SCFAs concentration more than 1 mM to provide constituent characteristics. In addition, solids collected from the primary sedimentation are introduced into the fermentor to increase COD and SCFAs concentration in the influent waste stream. The influent waste stream and a RAS are introduced into an aeration basin. The aeration basin is operated under HRT of approximately 1 day and F/M ratio of 0.8. A treated waste stream from the aeration basin is processed in a clarifier to separate AS from an effluent. A first portion of the AS is introduced into a return stream reactor. The return stream reactor is operated under HRT of 2.5 days and the F/M ratio of approximately 0.2. A second portion of the AS is separated by centrifugation followed by disinfection and an ensuing drying process to provide a stabilized PHA-laden biomass. Meanwhile, the effluent is ready to discharge. 
         [0030]    The present invention has been described in relation to particular embodiments, which are intended in all respects to be illustrative rather than restrictive. Alternative embodiments will become apparent to those of ordinary skill in the art to which the present invention pertains without departing from its scope. 
         [0031]    From the foregoing, it will be seen that this invention is one well adapted to attain all the ends and objects set forth above, together with other advantages which are obvious and inherent to the system and method. It will be understood that certain features and subcombinations are of utility and may be employed without reference to other features and subcombinations. This is contemplated by and is within the scope of the claims.