Abstract:
An apparatus that exposes a biofilm growth surface to liquid media as it rotates. A biofilm growth substratum is wound around a rotatable body in the form of a non-rigid material capable of supporting biofilm growth. A harvester receives the biofilm laden substratum, collects the biofilm as a biomass and reloads the substratum onto the rotatable body.

Description:
RELATED APPLICATIONS 
       [0001]    This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Patent Application No. 61/310,360 filed Mar. 4, 2010, and titled “Biomass production using a rotation bioreactor and spool harvester” which is incorporated herein by reference. 
     
    
     FIELD OF THE INVENTION 
       [0002]    This technology is an apparatus for growing and harvesting biomass for use as feedstock in, for example, the production of products or in wastewater remediation. 
       BACKGROUND 
       [0003]    Excess nitrogen, phosphorus, and other nutrients or compounds in discharged wastewaters can lead to downstream eutrophication and ecosystem damage. Advanced wastewater treatment technologies capable of removing these nutrients are expensive and often require the addition of chemical precipitants. Nitrogen and phosphorus can be removed naturally through biomass assimilation, but heterotrophic bacteria typically become carbon limited before removing all soluble N and P. Because microalgae are autotrophic, they can overcome this limitation and assimilate the remaining nutrients. In addition to the environmental benefits of harvesting algae grown during wastewater treatment, harvested microalgae are valuable as fertilizer, high-protein animal feed, and feedstock for the production of biofuels, including biodiesel and biomethane. Nutraceuticals, polymers, and other valuable products can be obtained from microalgae as well. Previously however, the realization of such benefits has been handicapped by an inability to find a reliable and cost effective apparatus and method of growing and harvesting the algae. 
         [0004]    Previous methods of growing algae at large scale include open outdoor pond systems and closed tubular photobioreactors. The most common outdoor pond design is the high rate algal pond, or raceway pond. These are shallow ponds that circle a volume of nutrient rich water by means of a paddle wheel. Although relatively inexpensive to build, large plots of land are required and the resulting algae yields are lower than with closed reactors. Tubular photobioreactors can often achieve higher cell concentrations than open ponds, but suffer from high material cost and frequent cell death due to inefficient gas exchange. Biofouling of the reactor walls also decreases light penetration and cleaning becomes an issue as well. With both methods, the resulting solution of suspended microalgae is very dilute, necessitating high cost methods of separation. 
         [0005]    Suspended microalgae must be removed from very dilute solutions and concentrated before further processing is possible. Current separation methods include filtration, sedimentation, centrifugation, dissolved air flotation, addition of electrolytes and polymers to induce coagulation and flocculation, and multiple combinations of these operations. Separation through filtration is difficult due to the small size of planktonic microalgae, and the sedimentation rate of algae is too slow for separation on a reasonable time scale. Dissolved air flotation requires high energy and high electrolyte and/or polymer addition to sufficiently flocculate microalgae. Centrifugation is currently the most common method used to separate algae from aqueous solutions; however, high upfront capital costs, power demand, and frequent maintenance make it uneconomical for large scale use. 
         [0006]    In addition to planktonic growth, microalgae are also capable of growing as biofilms attached to surfaces. Algal biofilms, or periphyton, are able to remove nutrients from wastewater just as suspended algae, and harvested biofilms can be processed into valuable products just as harvested suspended algae. When algae are grown as biofilms, the biomass is naturally concentrated and more easily harvested, leading to more direct removal and reduced downstream processing. The extracellular polymeric substance secreted by biofilms also increases the flocculation of associated suspended cells. Previously, however, there were no methods of growing and harvesting algal biofilms with any full scale potential. 
         [0007]    In addition to microalgae, other microorganisms are capable of growing as biofilms attached to surfaces. Biofilms are often complex mixed cultures containing microalgae, cyanobacteria, heterotrophic bacteria, nitrifying bacteria, microscopic fungi, and various combinations of these types of organisms. When grown as biofilms, the organism&#39;s morphology and metabolism are often different than when the organism is suspended. These changes are often beneficial, and can include increased production of a desired product. Biofilm reactors designed for the purpose of growing attached cultures include continuous stirred tank reactors (CSTR) with fibrous bed support, biofilm packed bed reactors (BPBR), biofilm trickling bed reactors (BTBR), and biofilm fluidized bed reactors (BFBR). Such reactors use a porous support or small granules as substrata for cell attachment and biofilm growth. These reactor configurations are often used to treat wastewater or produce a secreted product, but are limited in that harvesting of the biomass or intracellular product is not possible without high cost. 
       SUMMARY 
       [0008]    In one embodiment, we describe a reactor for the production of biomass involving a rotating cylinder or cylinders partially submerged in liquid media. The rotating cylinders are outfitted with a substratum capable of biofilm growth. The substratum is in a form that allows it to be wound around the cylinder, allowing the reactor to act as a spool and the harvesting of the biomass and reloading of the reactor are accomplished simultaneously. 
     
    
     
       BRIEF DESCRIPTION OF THE ILLUSTRATIONS 
         [0009]      FIG. 1  shows a rotating bioreactor partially submerged in liquid media with rope type substratum wound onto cylinder for biofilm growth. 
           [0010]      FIG. 2  shows a harvesting apparatus in conjunction with a rotating bioreactor. 
           [0011]      FIG. 3  shows a multiple cylinder setup. 
           [0012]      FIG. 4  shows a rotating reactor within a flotation frame. 
           [0013]      FIG. 5  shows a high rate algal pond with associated rotating bioreactors. 
           [0014]      FIG. 6  shows the photosynthetically active radiation cycle of bench scale reactors when operated at 4.8 rpm. 
           [0015]      FIG. 7  shows the growth curves of a suspended culture, initial biofilm culture, and secondary biofilm culture. 
           [0016]      FIG. 8  shows soluble P removal rates and soluble P concentrations of the suspended and biofilm reactors. 
           [0017]      FIG. 9  shows soluble N removal rates and soluble N concentrations of the suspended and biofilm reactors. 
       
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
       [0018]    In one embodiment, we describe a rotating bioreactor apparatus. In  FIG. 1  there is shown a body  10  partially submerged in a liquid medium  12 . In this embodiment the body is in the form of a right circular cylinder. Additional body formats may be utilized including, but not limited to, elliptic cylinder, parabolic cylinder, hyperbolic cylinder, generalized cylinder or oblique cylinder or any form with a rotational axis suitable for this purpose. 
         [0019]    One skilled in the relevant art will recognize that different formulations of liquid medium  12  will be used to produce different types of biomass. The liquid medium  12  may be a complex, defined, or selective growth medium. More specifically, the liquid medium  12  may be a complex medium including, but not limited to complex dextrose based media, sea water media, domestic wastewater, municipal wastewater, industrial wastewater, surface runoff wastewater, soil extract media, or any natural water containing detectable amounts of phosphorus or nitrogen; or a defined medium, including, but not limited to Bristol&#39;s medium, Bolds Basal medium, Walne medium, Guillard&#39;s f medium, Blue-Green medium, D medium, DYIY medium, Jaworski&#39;s medium, K medium, MBL medium, Jorgensen&#39;s medium, and MLA medium; or a selective medium including, but not limited to minimal media based on specific nutrient auxotrophy, and selective media that incorporates antibiotics. Depending on the chosen liquid medium  12  and seed culture, the resulting biofilm may be a mixed or pure culture and may be comprised of microalgae, cyanobacteria, nitrifying bacteria, heterotrophic bacteria, microscopic fungi, or any combination thereof. 
         [0020]    Still referring to  FIG. 1 , a rotation device  14  transmits rotational power to a drive shaft  16  that runs through the center of the cylinder  10  and is supported by a bearing  18  opposite the rotation device  14 . Where the drive shaft  16  enters and exits the cylinder  10 , a base plate  20  is used to connect the drive shaft  16  and the cylinder  10 . Holes  22  are made in the ends of the cylinder  10  to allow liquid media  12  to enter the cylinder  10 . A substratum  24  is placed around the cylinder  10  for biofilm growth. 
         [0021]    In more detail, still referring to  FIG. 1 , the rotation device  14  transmits rotational power to the drive shaft  16 , causing the cylinder  10  to rotate with the drive shaft  16 . As the cylinder  10  rotates, the biofilm substratum  24  placed on the surface of the cylinder  10  is alternately exposed to the liquid media  12  and the air. 
         [0022]    In further detail, still referring to  FIG. 1 , the biofilm substratum  24  may be in the form of a rope, cable or belt or the like such that it can be wound around the outer circumference of the cylinder  10 . The substratum  24  may be selected from a group comprising cotton, jute, hemp, manila, silk, linen, sisal, silica, acrylic, polyester, nylon, polypropylene, polyethylene, polytetrafluoroethylene, polymethylmethacrylate, polystyrene, polyvinyl chloride, or any other non-rigid material capable of supporting biofilm growth. One end of the substratum  24  is attached to one end of the surface of the cylinder  10  and wound around until the surface of the cylinder  10  may be sufficiently covered with the substratum  24 . The free end of the substratum  24  may then be attached to the surface of the cylinder  10  to keep the substratum  24  from unwinding during rotation of the cylinder  10 . 
         [0023]    In another embodiment, we describe a harvesting apparatus in conjunction with a rotating bioreactor. Referring now to  FIG. 2 , the biofilm is collected by detaching one end of the substratum  26  from the cylinder  28  and threading it through a scraper  30 . The scraper  30  may be a blade, series of blades, simple piece of rigid material with a hole in it, or more preferably, a unit with an adjustable diameter and/or constant tension settings like a hose clamp. The scraper  30  may be held in place by attachment to a support  32 . A reorientation system  34  is provided to prevent twisting or binding of the substratum  26 . The loose end of the substratum  26  is threaded through the scraper  30  and reorientation system  34  until it can be reattached to the cylinder  28 . As the cylinder  28  continues to rotate, the entire length of the substratum  26  may be pulled through the scraper  30  and pulley system  34  and rewound onto the cylinder  28 . To ensure the substratum  26  may be properly rewound onto the entire length of the cylinder  28 , the scraper  30 , support  32 , and pulley system  34 , are pulled on a support frame  36  along the length of the cylinder  28  at a rate such that the harvested portion of the substratum  26  may not be layered on top of itself as it is rewound. This may be accomplished with a lateral movement system  38  that may be powered by connection to the drive shaft  40  powering the cylinder  28 . Appropriate gear ratios may be chosen to achieve the desired pull rate and spacing of substratum  26 . As the biofilm is removed from the substratum  26 , it is gathered in a collection bin  42 . 
         [0024]    Referring now to another embodiment describing a multiple cylinder setup, shown in  FIG. 3 , a drive shaft  44  may be made long enough to support two or more cylinders  46  in a train formation. More cylinders  46  may be placed so that rotational power from a motor  48  is transferred to two or more drive shafts  44  through a power transfer mechanism like a roller chain  50 . The drive shafts  44  are supported by bearings  52  on each end. 
         [0025]    Referring to another embodiment shown in  FIG. 4 , the entire apparatus may be placed within a support frame  54  with attached floats  56 . The apparatus can then be placed at a suitable site and held in place using an anchor  58  or other suitable means of holding it in place. One application of this embodiment of the invention is a retrofitting of oxidation lagoons at a wastewater treatment plant. 
         [0026]    Referring to  FIG. 5 , another embodiment places the apparatus with a high rate algae pond  60  like a raceway or meandering ditch. The cylinders  62  may be rotated by the force of the passing water or powered by a motor and shaft connected to the cylinder. In a further embodiment, the cylinders  62  may be rotated by an air supply directed at the submerged perimeter of the cylinder in a direction perpendicular to the axis of rotation. In embodiments such as this, the biofilm enhances flocculation of the suspended culture, leading to inexpensive harvesting of all the biomass in the system. 
       EXAMPLES 
     Example 1 
       [0027]    In one embodiment, several bench scale units of the type shown in  FIG. 1  were used with 8 liters of chlorinated weak domestic strength wastewater as seeding media. A nested factorial experiment with triplicate replication of samples was established to determine the most suitable substrata for biofilm growth. The initial total suspended solids content of the wastewater was 42 mg/l. Concentrations of soluble phosphorus and nitrogen were brought to 5 mg/l and 36 mg/l respectively using KH 2 PO 4 , K 2 HPO 4 , and NaNO 3 . As a fed batch operation, N and P were added every 48 hours to give an average total P of 5.0 mg/l, and an average total N of 52.7 mg/l. Soluble N and P averaged 26.2 mg/l and 3.7 mg/l, respectively. A light cycle of 14 hours on, 10 hours off was used throughout the experiment.  FIG. 6  shows the cycle of photosynthetically active radiation (PAR) delivered to a point on the reactor during rotation at 4.8 rpm during periods while the lights were on. Biomass was harvested after 22 days of growth. This time included a recovery period following chlorination. Table 1 summarizes the results on the basis of mass per liquid surface area. 
         [0000]    
       
         
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Avg. Biomass yield of different substrate materials 
               
             
          
           
               
                   
                   
                 Avg. Biomass 
                   
               
               
                   
                   
                 Yield 
                 Std. 
               
               
                   
                 Substrata 
                 (g/m 2 ) 
                 Deviation 
               
               
                   
                   
               
             
          
           
               
                   
                 Cotton Rope 
                 91.2 
                 10.4 
               
               
                   
                 Cotton (High thread 
                 62.2 
                 0.9 
               
               
                   
                 count) 
                   
                   
               
               
                   
                 Jute 
                 51.4 
                 5.1 
               
               
                   
                 Cotton (Low thread 
                 51.3 
                 1.9 
               
               
                   
                 count) 
                   
                   
               
               
                   
                 Acrylic 
                 49.3 
                 0.4 
               
               
                   
                 Polyester 
                 19.3 
                 1.8 
               
               
                   
                 Polypropylene 
                 0 
                 0 
               
               
                   
                 Nylon 
                 0 
                 0 
               
               
                   
                 Construction Paper* 
                 0 
                 N/A 
               
               
                   
                 Sisal* 
                 0 
                 N/A 
               
               
                   
                 Lignin based cover* 
                 0 
                 N/A 
               
               
                   
                   
               
               
                   
                 *Materials showed some growth but biomass was not harvestable 
               
             
          
         
       
     
         [0028]    The substrata that were placed onto the cylinder as a sheet were harvested using a simple scraper blade. This proved to be difficult due to the constant adjustments required to scrape the uneven biofilm growth. Such substrata had also loosened during reactor operation causing frequent snagging and tearing against the scraper blade and rendering the materials unsuitable for future use. Cotton rope gave the highest biomass yields, and the rope construction allowed application of the harvesting method shown in  FIG. 2 . The cotton rope incurred no damage during harvesting and was immediately reused. 
       Example 2 
       [0029]    In another embodiment, the same procedure described in Example 1 was repeated with cotton rope as the only substratum. Triplicate samples were harvested after 10, 14, 18, 22, and 26 days of growth. Suspended cultures were also grown in reactor tanks of the same dimensions with the same light and nutrient conditions as the biofilm reactors. The same weak domestic strength wastewater was used to seed each type of reactor. Power input for mixing the suspended cultures was the same as the power input for rotating the cylinders. After each biofilm harvest, the substrata were reloaded onto the reactor to determine the secondary growth curve. Regrowth samples were harvested after 6, 10, 14, 18, and 22 days of growth. Growth in the suspended culture reactors was determined using the glass fiber filter method.  FIG. 7  shows the growth curves of the initial biofilms, secondary biofilms, and suspended cultures. It can be seen that the biofilm grows at a much faster rate after the initial harvest. This is most likely due to the residual biomass left on the substratum after scraping. This secondary growth curve represents the true productivity of the reactor when operated continuously. Table 2 compares the maximum productivity obtained by each type of growth and at what time it was obtained. 
         [0000]    
       
         
               
             
               
               
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 Maximum productivity obtained by different growth types 
               
             
          
           
               
                   
                   
                 Yield* 
                 Time 
                 Productivity* 
               
               
                   
                 Growth Type 
                 (g/m 2 ) 
                 (days) 
                 (g/m 2  day) 
               
               
                   
                   
               
               
                   
                 Biofilm initial 
                 51.6 ± 6.6 
                 22 
                 2.4 ± 0.3 
               
               
                   
                 Biofilm 
                 98.9 ± 9.3 
                 18 
                 5.5 ± 0.5 
               
               
                   
                 regrowth 
                   
                   
                   
               
               
                   
                 Suspended 
                 20.4 ± 1.4 
                 22 
                 0.9 ± 0.1 
               
               
                   
                   
               
               
                   
                 *plus and minus one standard deviation from the mean 
               
             
          
         
       
     
       Example 3 
       [0030]    In another embodiment, nitrogen and phosphorus concentration data from the experiment of Example 2 were analyzed to determine the wastewater remediation ability of the suspended culture and the biofilms. After filtration of wastewater samples, soluble N concentrations were determined using the chromotropic acid method for nitrate-N and the salicylate method for ammonia-N. Soluble P as orthophosphate was determined using the ascorbic acid method. The wastewater samples were also analyzed for total N and P using the chromotropic acid method with alkaline persulfate digestion and the molybdovanadate method with acid persulfate digestion, respectively. 
         [0031]      FIG. 8  shows soluble P removal rates for the suspended and biofilm cultures.  FIG. 9  shows soluble N removal rates for the suspended and biofilm cultures. It can be seen that the biofilm reactors demonstrated higher removal of both nitrogen and phosphorus compared to the suspended culture reactors. Furthermore, these nutrients could be easily removed from the system by simply removing the biofilm as shown in  FIG. 2 , whereas the suspended cultures would have to be removed through centrifugation, filtration, or the like to completely remove the nutrients from the system. 
       Example 4 
       [0032]    In another embodiment, as the biofilms of the experiments of Example 1 and Example 2 were grown, a visual observation of the wastewater turbidity was made for each tank containing a rotating bioreactor. It was observed that at some point during operation, typically between 12-18 days of growth, the suspended microorganisms in the wastewater associated with the rotating bioreactors underwent spontaneous autoflocculation and settled to the bottom or floated to the top of the reactor tank. Such flocculated biomass would be much easier to harvest than a suspended culture. 
         [0033]    From the foregoing description, one skilled in the art can easily ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make changes and modifications of the invention to adapt it to various usage and conditions. The preceding preferred specific embodiments are to be construed as merely illustrative, and not limiting of the scope of the invention in any way whatsoever.