WO1995015924A1 - Composition and method for sewage treatment using fungal and bacterial enzymes - Google Patents
Composition and method for sewage treatment using fungal and bacterial enzymes Download PDFInfo
- Publication number
- WO1995015924A1 WO1995015924A1 PCT/US1994/013520 US9413520W WO9515924A1 WO 1995015924 A1 WO1995015924 A1 WO 1995015924A1 US 9413520 W US9413520 W US 9413520W WO 9515924 A1 WO9515924 A1 WO 9515924A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- bacillus
- composition
- cellulase
- composition according
- cultures
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/342—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the enzymes used
Definitions
- This invention relates to cellulose degradation by fungal and bacterial enzymes.
- the combination of enzymes is useful for sewage treatment, particularly in septic tanks.
- sewage treatment centers may employ microorganisms for the degradation of sewage.
- sewage contains water, organic waste (containing carbohydrates, fats and proteins), and cellulose from paper products.
- Cellulose may represent up to about 15% of the solids in raw (untreated) sewage.
- the organic, non-cellulosic waste component of sewage is more easily degraded than the cellulose component.
- Carbohydrates, fats and proteins making up the organic waste are fairly easily digested by extracellular enzymes released outside the cell of selected bacteria. The degradation of cellulose, however, remains a problem in many forms of sewage treatment.
- the degradation of cellulose to glucose is a stepwise process.
- cellulose is hydrolyzed by the action of an endoglucanase that breaks bonds along the amorphous regions of cellulose.
- This enzymatic reaction carries out the cleavage of the beta (1 > 4) bonds producing cellobiose which will be removed from the nonreducing ends of the molecule by the action of a beta (1 > 4) exoglucanase.
- the cellobiose is hydrolyzed by a beta (1 > 4) glucosidase to glucose.
- the breakdown of cellulose to glucose involves a complex of enzymes. Sufficient amounts of these enzy es are not believed to be produced by natural bacteria. The lack of sufficient enzymatic activity is particularly evident in septic tanks, where cellulose sediment is a problem. New methods to reduce the cellulose sediment in raw sewage are needed.
- a sewage treatment composition comprising Bacillus spp. cultures in combination with fungal cellulase.
- the combination of the extracellular enzymes produced by bacteria cultures from Bacillus spp. and fungal cellulase results in a synergistic degradation of cellulose.
- Results show a significant enhancement in the production of glucose as a result of cellulose degradation when sewage containing cellulose is contacted with the inventive composition.
- the composition is a broad based system capable of breaking down carbohydrates, fats and proteins in addition to enhanced cellulose degradation. Because the composition contains enzymes from naturally occurring microorganisms, it is particularly useful as a septic tank additive.
- the invention also provides a novel method for using the Bacillus spp. cultures and fungal cellulase to degrade carbohydrates, protein, fat, and cellulose, and mixtures thereof.
- Bacillus spp. are known naturally occurring bacteria as identified on pages 1105 to 1139 of the eighth Edition of Bergey ' ⁇ Manual of Determinative Bacteriology, published by The Williams and Wilkins Co., 1986.
- Preferred Bacillus species include B. subtili ⁇ , B . licheni formi ⁇ , B . mega teri um, and mixtures thereof. More preferably, the bacteria culture is a mixture of B . ⁇ ubtilis, B . li cheni formi ⁇ , and B. megaterium.
- bacteria cultures may be prepared as spores to extend the period that the cultures may be stored.
- the Bacillus spp. cultures are present in the composition as spores.
- the spores become enzyme producing organisms when exposed to nutrients such as sewage.
- the spores When exposed to sewage the spores generate into bacteria producing extracellular enzymes that are particularly effective in degrading carbohydrates, fats and proteins.
- the spore count of Bacillus spp. employed in the composition may vary greatly, depending upon the type of sewage to be treated, the size of the sewage treatment facility, the frequency of treatment of the sewage with the composition, and so on.
- the active ingredient portion of the composition is defined as the bacteria culture and fungal enzyme.
- a concentration range of bacteria cultures for a composition prepared as a typical septic tank additive preferably employs at least about 10 ⁇ spores/g of composition (active ingredient) , with the upper limit concentration of spores generally limited only by cost. More preferably at least 10 ⁇ spores/g and most preferably from 10 ⁇ to 10 ⁇ spores/g of composition (active ingredient) is employed in the composition.
- the cellulase is isolated from A ⁇ pergillu ⁇ niger fungus.
- the enzyme may be extracted from the fungal culture by any known means, and is widely available commercially from, for example, Novo Nordisk, Ct.; Sigma Chemical, St. Louis, Mo.; and George A.
- the fungus is aerobic, and sewage treatment is largely in a submerged anaerobic environment, it is preferred that the cellulase enzyme is separated from the fungus as employed in this invention.
- the Bacillus spp. bacteria are facultative anaerobic and thus thrive in the typically anaerobic conditions of sewage treatment.
- the specific activity and amount of the fungal cellulase enzyme employed in the composition is widely variable and may be adjusted according to the enzymatic needs of the system employing the composition. For example, with waste systems having a particularly high content of cellulose, large amounts of the cellulosic enzymes would be preferred.
- the activity of the fungal enzyme employed is preferably at least about 1000 CU/g of active ingredient portion of composition (with the upper limit of concentration of enzyme generally limited only by cost) .
- the enzyme range is more preferably from 1500 to 2500 CU/g and most preferably from 1500 to 2000 CU/g of active ingredient portion of composition.
- the ratio of bacteria culture to fungal enzyme may vary greatly. Preferably the ratio is anywhere between about 10:90 to about 99.99:0.01 percent by weight of active ingredient bacteria culture:fungal enzyme. As known to those skilled in the art, the ratio may be adjusted depending upon the type of material to be treated, the spore count and specific activity of raw materials, and so on.
- the composition may also include optional fillers and additives to facilitate storage or delivery of the spores and fungal enzyme into the treatment facility.
- Fillers that may be used include, but are by no means limited to, alkali metal salts (such as NaCl, NaS04, CaC ⁇ 3, mixtures thereof and so on), inert preparations (such as milorganite) , mixtures thereof, and so on.
- the composition may be prepared as a liquid or powder by any means known to those skilled in the art.
- the enzymes utilized in the inventive composition are produced by naturally occurring organisms.
- the composition is useful for many industrial applications where broad based degradation of components typical of sewage (e.g. carbohydrates, proteins, fats and cellulose) is desired.
- the combination of Bacillus spp. enzymes and the fungal cellulase has been found to be synergistic.
- a smaller amount of bacteria and fungal enzymes used in combination was found to be more effective in degrading cellulose than when a larger amount of plain fungal enzyme was used.
- the inventive combination offers a broad based sewage treatment system as well as a means of producing glucose from cellulose, particularly useful in industrial applications where cellulose is a waste product.
- the enzymatic action of the inventive composition may occur over a wide pH range.
- the pH range of the media to be treated falls within about 4 to about 10, with more preferably the pH having a value between 6 and 8.
- the temperature range of the media to be treated may vary greatly, although optimum enzymatic action preferably occurs within a temperature range of from about 10°C to about 45°C and more preferably between 20°C and 35°C.
- Degradation of cellulose may also occur with enzymes separated from the Bacillus spp. and combined with cellulase of a fungal origin (separated or unseparated from the fungus).
- the fungus is an aerobic microorganism and the Bacillus spp. a facultative anaerobic microorganism, thus the oxygen content of the substrate environment must be considered in preparing the composition.
- the dosage, frequency of use, as well as the concentration of the active ingredient portion of the composition are interdependent variables that will also vary widely depending upon the environment to be treated, the concentration of particles to be degraded, prior usage of microorganisms, and so on. Adjustments to these variables may be accomplished by routine procedures known to those skilled in the art.
- an effective amount of the active ingredient portion of the composition is at least about 10 g, more preferably at least about 100 g (with the upper limit of the amount used limited primarily by cost), and most preferably from 150 g to 1000 g.
- compositions described in the examples used spores isolated from Bacill us subtili ⁇ , Bacillu ⁇ licheniformi ⁇ , and Bacillus mega terium, and fungal cellulase isolated from A ⁇ pergillu ⁇ niger. Both the spores and cellulase were obtained from the George A. Jeffreys Company. The culture has a count of 10° spores/gram of active ingredient portion of the composition. The cellulase had a specific activity of 1600 CU/g of active ingredient portion of the composition. (As obtained from supplier, actual cellulase enzyme activity was approximately 128,000 CU/g.) The milorganite was purchased from Milwaukee Metropolitan Sewage District, Milwaukee, WI.
- Examples 1 and 2 and Comparative Examples 1 and 2 Synthetic sewage was prepared with 5% protein, 5% fat, 5% cellulose, and the remainder distilled water. The synthetic waste was placed in a 35 ml test tube for each composition tested.
- Example 1 Composition A was diluted 10% with the waste for a final cellulase concentration of 0.050% (64 CU/g) . After 48 h glucose production (thus indicating the level of cellulose degradation) was measured as 4.83 g/lt using the DNS method, as recorded in Table I below.
- Example 2 Example 1 was repeated with the exception that Composition A was diluted 2.5% with waste for a final cellulase concentration of 0.0125% (16 CU/g). After 47 h glucose production was measured by DNS as 1.31 g/lt, as recorded in Table I below. Comoarative Example 1 Example 1 was repeated with the exception that only cellulase was added to the synthetic waste at a concentration of 0.5% (640 CU/g). After 48 h glucose production was measured by DNS as 1.41 g/lt, as recorded in Table I.
- Example 1 was repeated with the exception that Composition A did not have any fungal cellulase and CaC ⁇ 3 and NaS ⁇ 4 were replaced with an equivalent amount milorganite. After 48 h glucose production was measured by DNS as 0.17 g/lt, as recorded in Table I. Table 1 summarizes data obtained in Examples 1 and 2 and Comparative Examples 1 and 2. When the fungal cellulase was added to the culture a significant increase in the production of glucose was detected. As Examples 1 and 2 show more, or similar amount of glucose was produced by the composition, once the cellulase was added, than with the enzyme by itself though the cellulase concentration in Examples 1 and 2 was approximately ten times less than with the enzyme by itself. Therefore, these major differences between the enzymes by itself and the combination of fungal and bacterial enzymes were due to the synergistic effect of both bacterial and fungal enzymes. Similar results were found using raw sewage (Table 2).
- Example 3 Raw sewage (obtained from the Ridgewood waste water treatment plant, Ridgewood, N.J.) was placed into a 35 ml tube and Composition B was diluted 2.5% for a final cellulase concentration of 0.0125% (16 CU/g). After 48 h glucose production (thus indicating the level of cellulose degradation) was measured as 0.291 g/lt using the DNS method, as recorded in Table 2 below.
- Example 3 was repeated with the exception that only cellulase was added to the sewage at a concentration of 0.5% (640 CU/g). After 48 h glucose production was measured by DNS as 0.128 g/lt, as recorded in Table 2.
- Example 3 was repeated with the exception that Composition B did not have any fungal cellulase and CaC ⁇ 3 and NaS04 were replaced with milorganite. After 48 h glucose production was measured by DNS as 0.029 g/lt, as recorded in Table 2.
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- Life Sciences & Earth Sciences (AREA)
- Microbiology (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Hydrology & Water Resources (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Treatment Of Sludge (AREA)
- Detergent Compositions (AREA)
Abstract
Description
Claims
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP95904120A EP0733025A1 (en) | 1993-12-09 | 1994-11-21 | Composition and method for sewage treatment using fungal and bacterial enzymes |
NZ277630A NZ277630A (en) | 1993-12-09 | 1994-11-21 | Composition and method for sewage treatment using fungal and bacterial enzymes |
JP7516207A JPH09509308A (en) | 1993-12-09 | 1994-11-21 | Compositions and methods for treating sewage using fungal and bacterial enzymes |
BR9408267A BR9408267A (en) | 1993-12-09 | 1994-11-21 | Process composition to use process composition to produce glycols from cellulose process to degrade sewage and septic tank additive |
AU12931/95A AU682565B2 (en) | 1993-12-09 | 1994-11-21 | Composition and method for sewage treatment using fungal and bacterial enzymes |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US16460993A | 1993-12-09 | 1993-12-09 | |
US164,609 | 1993-12-09 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1995015924A1 true WO1995015924A1 (en) | 1995-06-15 |
Family
ID=22595281
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1994/013520 WO1995015924A1 (en) | 1993-12-09 | 1994-11-21 | Composition and method for sewage treatment using fungal and bacterial enzymes |
Country Status (10)
Country | Link |
---|---|
EP (1) | EP0733025A1 (en) |
JP (1) | JPH09509308A (en) |
CN (1) | CN1147803A (en) |
AU (1) | AU682565B2 (en) |
BR (1) | BR9408267A (en) |
CA (1) | CA2178344A1 (en) |
NZ (1) | NZ277630A (en) |
SG (1) | SG52242A1 (en) |
WO (1) | WO1995015924A1 (en) |
ZA (1) | ZA949839B (en) |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1997044281A1 (en) * | 1996-05-21 | 1997-11-27 | Bevil S.P.R.L. | Biological purification of septic tanks by extender effect |
US5812602A (en) * | 1996-11-14 | 1998-09-22 | Motorola Inc. | System and device for, and method of, communicating according to a trellis code of baseband signals chosen from a fixed set of baseband signal points |
US5822371A (en) * | 1997-02-14 | 1998-10-13 | General Datacomm Inc. | Mapper for high data rate signalling |
US5838724A (en) * | 1997-02-14 | 1998-11-17 | General Datacomm, Inc. | Spectral and power shaping mapper for high data rate signalling |
US5862179A (en) * | 1997-02-14 | 1999-01-19 | General Datacomm, Inc. | Mapper for high data rate signalling |
US5875229A (en) * | 1996-10-15 | 1999-02-23 | Motorola Inc. | System and device for, and method of, detecting, characterizing, and mitigating deterministic distortion in a communications network |
US6185249B1 (en) * | 1999-01-28 | 2001-02-06 | Ic Tel Inc. | Translation table design for a PCM modem |
US6560277B2 (en) | 2001-02-09 | 2003-05-06 | Pc Tel, Inc. | Distinguishing between final coding of received signals in a PCM modem |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2319504A1 (en) | 2009-11-07 | 2011-05-11 | Laboratorios Del. Dr. Esteve, S.A. | Pharmaceutical solid dosage form |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2482130A1 (en) * | 1980-05-12 | 1981-11-13 | Lumer Ste Indle Produits Chimi | Inoculum for septic tanks and purificn. digesters - prepd. by drying bacterial cultures with molecular sieve, then adding enzyme and dispersant |
GB2102428A (en) * | 1981-07-16 | 1983-02-02 | Unisearch Ltd | Enzymatic hydrolysis of cellulosic material |
US4940539A (en) * | 1989-05-08 | 1990-07-10 | Semco Laboratories, Inc. | Grease trap construction |
WO1993005187A1 (en) * | 1991-08-30 | 1993-03-18 | United Laboratories, Inc. | Method of separating oleophilic-hydrophobic material from wash water |
FR2699525A1 (en) * | 1992-12-22 | 1994-06-24 | Hecke Jean Claude Van Den | Compsn. for purifying contaminated aq. liquids pref. water |
-
1994
- 1994-11-21 WO PCT/US1994/013520 patent/WO1995015924A1/en not_active Application Discontinuation
- 1994-11-21 AU AU12931/95A patent/AU682565B2/en not_active Ceased
- 1994-11-21 NZ NZ277630A patent/NZ277630A/en unknown
- 1994-11-21 BR BR9408267A patent/BR9408267A/en not_active Application Discontinuation
- 1994-11-21 JP JP7516207A patent/JPH09509308A/en active Pending
- 1994-11-21 CN CN94194795A patent/CN1147803A/en active Pending
- 1994-11-21 SG SG1996001168A patent/SG52242A1/en unknown
- 1994-11-21 EP EP95904120A patent/EP0733025A1/en not_active Withdrawn
- 1994-11-21 CA CA002178344A patent/CA2178344A1/en not_active Abandoned
- 1994-12-09 ZA ZA949839A patent/ZA949839B/en unknown
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2482130A1 (en) * | 1980-05-12 | 1981-11-13 | Lumer Ste Indle Produits Chimi | Inoculum for septic tanks and purificn. digesters - prepd. by drying bacterial cultures with molecular sieve, then adding enzyme and dispersant |
GB2102428A (en) * | 1981-07-16 | 1983-02-02 | Unisearch Ltd | Enzymatic hydrolysis of cellulosic material |
US4940539A (en) * | 1989-05-08 | 1990-07-10 | Semco Laboratories, Inc. | Grease trap construction |
WO1993005187A1 (en) * | 1991-08-30 | 1993-03-18 | United Laboratories, Inc. | Method of separating oleophilic-hydrophobic material from wash water |
FR2699525A1 (en) * | 1992-12-22 | 1994-06-24 | Hecke Jean Claude Van Den | Compsn. for purifying contaminated aq. liquids pref. water |
Cited By (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1997044281A1 (en) * | 1996-05-21 | 1997-11-27 | Bevil S.P.R.L. | Biological purification of septic tanks by extender effect |
US5875229A (en) * | 1996-10-15 | 1999-02-23 | Motorola Inc. | System and device for, and method of, detecting, characterizing, and mitigating deterministic distortion in a communications network |
US5812602A (en) * | 1996-11-14 | 1998-09-22 | Motorola Inc. | System and device for, and method of, communicating according to a trellis code of baseband signals chosen from a fixed set of baseband signal points |
US5822371A (en) * | 1997-02-14 | 1998-10-13 | General Datacomm Inc. | Mapper for high data rate signalling |
US5838724A (en) * | 1997-02-14 | 1998-11-17 | General Datacomm, Inc. | Spectral and power shaping mapper for high data rate signalling |
US5862179A (en) * | 1997-02-14 | 1999-01-19 | General Datacomm, Inc. | Mapper for high data rate signalling |
US6115415A (en) * | 1997-02-14 | 2000-09-05 | General Data Comminc. | Mapper for high data rate signalling |
US6185249B1 (en) * | 1999-01-28 | 2001-02-06 | Ic Tel Inc. | Translation table design for a PCM modem |
US6560277B2 (en) | 2001-02-09 | 2003-05-06 | Pc Tel, Inc. | Distinguishing between final coding of received signals in a PCM modem |
US6778597B2 (en) | 2001-02-09 | 2004-08-17 | Pctel, Inc. | Distinguishing between final coding of received signals in a PCM modem |
Also Published As
Publication number | Publication date |
---|---|
SG52242A1 (en) | 1998-09-28 |
BR9408267A (en) | 1996-12-10 |
CA2178344A1 (en) | 1995-06-15 |
ZA949839B (en) | 1996-06-10 |
NZ277630A (en) | 1997-10-24 |
AU682565B2 (en) | 1997-10-09 |
EP0733025A1 (en) | 1996-09-25 |
AU1293195A (en) | 1995-06-27 |
CN1147803A (en) | 1997-04-16 |
JPH09509308A (en) | 1997-09-22 |
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