KR20010100334A - Microbial blends for wastewater treatment and its manufacturing method - Google Patents

Microbial blends for wastewater treatment and its manufacturing method Download PDF

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KR20010100334A
KR20010100334A KR1020000022425A KR20000022425A KR20010100334A KR 20010100334 A KR20010100334 A KR 20010100334A KR 1020000022425 A KR1020000022425 A KR 1020000022425A KR 20000022425 A KR20000022425 A KR 20000022425A KR 20010100334 A KR20010100334 A KR 20010100334A
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wastewater
wastewater treatment
spawn
liquid
preparing
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오덕환
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양갑석
주식회사 고합
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • 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
    • C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
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    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus
    • C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/425—Serratia

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Abstract

유기성 폐수의 정화를 위한 생물학적 처리에 있어 중요한 역할을 하는 유용 미생물군을 유통기간 중 생존율 및 그 활성을 높은 상태로 유지토록 하는 액상 미생물제제의 제조방법Method for producing a liquid microbial agent to maintain a useful survival rate and activity of the microbial group that plays an important role in biological treatment for the purification of organic wastewater

Description

폐수 및 오수처리용 미생물 제제 및 그 제조방법 {Microbial blends for wastewater treatment and its manufacturing method}Microbial blends for wastewater treatment and its manufacturing method

산업의 급속한 발달과 인구의 기하 급수적인 증가로 인한 환경오염의 문제는 인류의 생존을 위협하는 정도에까지 발전되었다. 이러한 환경오염의 주된 원인중의 하나인 산업폐수 및 도시하수 등은 우리 인간의 중요한 식수원인 하천과 지하수를 극심하게 오염시키므로 우리 국민의 보건을 위협하고 있음은 주지의 사실이다.The problem of environmental pollution due to the rapid development of industry and the exponential increase of population has developed to the extent that it threatens human survival. It is well known that industrial wastewater and urban sewage, which are one of the main causes of environmental pollution, severely pollute rivers and groundwater, which are important drinking water sources for humans, and thus threaten the health of our people.

일반적으로 폐수처리 기술은 물리적 처리, 화학적 처리 및 생물학적 처리의 세 가지로 대변할 수 있으나 그 중에서도 유용 미생물에 의한 생물학적 처리법은 일반적인 자연 생태계의 자정작용을 특정 설비 내에 인위적으로 조성, 일정 조건아래서 제어·관리하는 것으로, 유기성 폐수(도시하수 및 식품, 제지폐수 등)의 처리법으로는 가장 바람직한 것이 바로 생물학적 처리이다. 생물학적 폐수처리에 관여하는 미생물은 BOD와 COD의 1차 분해자 역할을 하는 박테리아(Bacteria)에서부터 곰팡이류(Fungi), 조류(Algae), 원생동물(Protozoa) 등이 있다. 이들 중에서 폐수처리에핵심적인 역할을 하는 것이 박테리아와 원생동물이다. 박테리아는 크기가 일반적으로 0.8∼5㎛ 정도이며, 영양소에 따라 유기물 탄소원과 에너지원으로 이용하는 종속 영양세균과 무기물을 이용하는 독립영양세균으로 나누어진다. 생물학적 폐수처리에는 주로 생장에 유기물을 이용하는 종속 영양세균이 이용된다. 원생동물은 다양한 형태로 생물학적 폐수처리 마지막 단계에 출현하여 BOD 또는 COD 원인물질을 분해, 증식한 박테리아의 최종 포식자가 되며 폐수처리의 지표 미생물로 이용되지만 박테리아보다 주위 환경변화에 민감하여 독성물질이나 악성 산업폐수에 출현하는 경우는 다소 드물다.In general, wastewater treatment technology can be represented by three kinds of physical treatment, chemical treatment, and biological treatment. Among them, biological treatment by useful microorganisms artificially controls the self-cleaning of general natural ecosystems in a specific facility, under certain conditions. In terms of management, biological treatment is the most preferable method for treating organic wastewater (urban wastewater and food, paper wastewater, etc.). Microorganisms involved in biological wastewater treatment include bacteria (Bacteria), which act as primary decomposers of BOD and COD, fungi, algae, and protozoa. Among them, bacteria and protozoa play a key role in wastewater treatment. Bacteria generally have a size of 0.8 ~ 5㎛, and depending on nutrients, they are divided into heterotrophic bacteria used as organic carbon sources and energy sources, and autotrophic bacteria using inorganic materials. Heterotrophic bacteria are used mainly for biological wastewater treatment using organic materials for growth. Protozoa appear in various forms at the end of biological wastewater treatment and become the final predators of bacteria that decompose and propagate BOD or COD-causing substances and are used as indicator microorganisms in wastewater treatment, but are more sensitive to changes in the surrounding environment than bacteria. It rarely appears in industrial wastewater.

생물학적 폐수처리 현장에서는 폭기조내의 박테리아와 원생동물의 상태를 적절히 유지하는 것이 필수적이다. 폐수처리 시설이 아무리 합리적으로 설계되고 최신 설비를 갖추었을지라도 그 시설의 조작 관리는 폭기조내 유용 미생물군의 증식대사를 적정한 수준으로 유지하도록 조정을 하는 것이 폐수처리 기술의 핵심인 것이다.At the biological wastewater treatment site, it is essential to maintain the proper condition of bacteria and protozoa in the aeration tank. No matter how reasonably designed and state-of-the-art the facility is, wastewater treatment is at the heart of wastewater treatment technology, in which the control of its operation is controlled to maintain the proper growth rate of useful microbial populations in the aeration tank.

본 발명은 유기성 폐수의 정화를 위한 생물학적 처리에서 중요한 역할을 하는 유용 미생물군을 유통기간 중 생존율 및 그 활성을 높은 상태로 유지토록 하는 액상 미생물 제제의 제조방법에 관한 것이다. 특히 본 발명은 액상 미생물 제제를 유기성 폐수의 생물학적 폐수처리 시스템에 적용함으로써 폭기조내 유용 미생물군의 생육을 촉진하고 폐수처리 시스템의 활성을 지속적으로 유지하여 유해 미생물의 발생을 억제함과 동시에 팽화, 벌킹(Bulking) 및 쇼크(Shock)의 현상을 방지하여 효과적인 폐수처리를 이루고 자 하는 것이다.The present invention relates to a method for producing a liquid microbial preparation for maintaining a high survival rate and activity of the useful microbial group in the biological treatment for the purification of organic wastewater during the distribution period. In particular, the present invention promotes the growth of useful microbial groups in the aeration tank by applying the liquid microbial agent to the biological wastewater treatment system of organic wastewater and maintains the activity of the wastewater treatment system to suppress the generation of harmful microorganisms and at the same time, swelling and bulking It is to achieve effective wastewater treatment by preventing the phenomenon of bulking and shock.

아무리 뛰어난 효능을 지닌 균주를 사용할 지라도 정상적인 상태로 유통될 경우 자가 소화되어 생존율이 매우 낮아지게 되어 그 효능을 발휘할 수 없게되는데 일반적으로 시중에 나와있는 액상 종균제들은 다른 미생물들에 의해 쉽게 오염될 수 있는 용기에서 균을 적정 수준의 농도까지 배양치 못한 채 배양액 자체를 포장용기에 담아 유통시켜 종종 문제를 야기하며 심한 구린내 및 악취를 유발하고 때로는 규격에 표시된 생균수에 훨씬 못 미치는 제품을 저가로 내어 시장을 교란시킨다. 심한 경우는 수도물 내지 지하수 자체를 종균제인 양 일반 오수·정화조의 처리장에 납품하기도 한다.No matter how good strains are used, if they are distributed under normal conditions, they will be self-digested and their survival rate will be very low, so they will not be able to exert their efficacy. Generally, the liquid spawns on the market can be easily contaminated by other microorganisms. It is often a problem to circulate the culture solution itself in a packaging container without incubating the germs to an appropriate level in a container. Disrupt the market. In severe cases, tap water or groundwater itself may be delivered to the treatment plant of a general sewage / septic tank.

유기성 폐수의 경우 그 내용물은 단백질, 탄수화물, 및 지질이 주된 구성요인이며 이러한 것들을 분해하기 위해서 각각의 것들마다 독특한 효소(Enzyme)가 작용하게 됨은 주지의 사실이다. 자연계에 존재하는 미생물은 분비할 수 있는 효소의 종류 및 능력이 각기 다르나 이것 모두 분해할 수 있는 균주가 많이 존재하며 우수한 균주들이 널리 알려져 있다. 이러한 균주가 폐수처리 시스템 내 폭기조에서 우점적으로 증식한다면 폐수처리의 효과는 높을 것이다. 본 발명에서는 3가지 효소 분비능을 시험하여 3가지 효소를 모두 분비하고 분비능이 우수한 균주를 선별하여 액상 미생물 제제의 주된 요소로 삼았다. 대표적인 균주로Serratia,B. subtilis및Rhodopseudomonas를 들 수 있다. 이 균주들은 위에서 언급한 3가지 효소 외에도 다른 유기물이나 화합물을 분해하는 효소(lipase, xylanase etc.)들도 우수하게 분비하는 것으로 판명되었다.Serratia는 현재 효소 생산균주, 색소 생성균주, 의약용 약품 생산균주로 광범위하게 활용되고 있으나Serratia의 색소는 다른 미생물에게 독성을 지니는 바, 본 액상 종균제에 첨가되는 균주는 색소 생성능을 돌연변이에 의해 제거시켜 다른 미생물에게 전혀 해를 끼치지 않도록 했다.B. subtilis는 중온뿐만 아니라 고온에서조차 높은 활성을 지니는 것으로 판명되었으며 높은 NaCl농도(3∼5)에서도 잘 자라는 것으로 평가되었다. 광합성 세균은 혐기적 및 호기적 조건에서도 잘 자라는 통성 혐기성 균주로 고농도 유기물 및 난분해성 Hydrocarbon을 잘 분해하는 것으로 드러났다. 이들 균주의 드러난 특징들은 어느 환경에서든지 잘 적응하여 높은 활성을 나타낸다는 점에서 폐수처리 시스템에서 유리한 점을 지닌다. 본 발명의 또 다른 특징은 균의 성장을 억제하는 안정제 Glutaraldehyde 및 침전물의 생성을 억제하는 천연 점증물질인 알긴산을 사용하여 유통중의 안정성을 확보한 점이다.In the case of organic wastewater, its contents are the major constituents of proteins, carbohydrates, and lipids, and it is well known that each enzyme has a unique enzyme to break down these substances. Microorganisms present in nature are different in type and ability to secrete enzymes, but there are many strains capable of degrading all of them and excellent strains are widely known. If these strains proliferate predominantly in the aeration tank in the wastewater treatment system, the effect of the wastewater treatment will be high. In the present invention, three enzyme secretion ability was tested and all three enzymes were secreted, and a strain having excellent secretion ability was selected as the main element of the liquid microbial preparation. Representative strains include Serratia , B. subtilis and Rhodopseudomonas . In addition to the three enzymes mentioned above, these strains have been shown to secrete other enzymes (lipases, xylanase etc.) that degrade other organisms or compounds. Serratia is currently widely used as an enzyme-producing, pigment-producing, and pharmaceutical drug-producing strain. However, Serratia 's pigments are toxic to other microorganisms. So no harm to other microorganisms. B. subtilis was found to have high activity at high as well as mesophilic temperatures, and was well grown at high NaCl concentrations (3 to 5). Photosynthetic bacteria are well-developed anaerobic strains that grow well under anaerobic and aerobic conditions, and have been shown to degrade high concentrations of organics and hardly degradable hydrocarbons. The revealed features of these strains are advantageous in wastewater treatment systems in that they adapt well in any environment and exhibit high activity. Another feature of the present invention is to secure stability in circulation by using a stabilizer Glutaraldehyde that inhibits the growth of bacteria and alginic acid which is a natural thickener that suppresses the production of precipitates.

본 발명의 제품은 스팀에 의해 내부 배양액을 멸균하고 강제 통기에 의한 산소 공급과 Heater로서 적정 온도를 유지하여 각각의 균주가 고농도로 자라도록 한다. 유기성 폐수처리를 위한 본 발명의 제조공정에 대해 구체적이고 상세하게 설명하면 다음과 같다.The product of the present invention sterilizes the internal culture by steam and maintains the optimum temperature as the oxygen supply and the heater by forced aeration so that each strain grows at a high concentration. Hereinafter, the manufacturing process of the present invention for organic wastewater treatment will be described in detail.

제 1공정 : Seed 배양과정Step 1: Seed Culture

준비된 Plate나 글리세롤을 이용하여 보관된 균주를 고온고압 살균한 영양배지에서30℃, 180rpm의 조건으로 20시간 배양한다.Incubate the stored strain using the prepared plate or glycerol for 20 hours at 30 ℃, 180rpm in a nutrient medium sterilized at high temperature and high pressure.

제 2공정 : 생산배지의 스팀 멸균과정Second Process: Steam Sterilization of Production Medium

배지조성(리터 기준)Medium composition (based on liters)

포도당 ---------- 20gGlucose ---------- 20g

효모 엑기스 ------ 3gYeast Extract ------ 3 g

구연산 ----------- 1gCitric Acid ----------- 1g

황산암모늄 ------- 1gAmmonium Sulfate ------- 1g

일인산카리 ------- 1gMonophosphate ------- 1 g

이인산소다 ------- 1gSodium Diphosphate ------- 1g

황산마그네슘 ----- 0.2gMagnesium sulfate ----- 0.2g

Corn Steep Liquor -- 1㎖Corn Steep Liquor-1 ml

미네랄 용액 ------- 1㎖Mineral solution ------- 1ml

준비된 배지를 산도 7.5로 조정한 후, 121℃에서 15분 스팀멸균처리After adjusting the prepared medium to pH 7.5, steam sterilization for 15 minutes at 121 ℃

제 3공정 : 균체량 최대생산을 위한 배양과정3rd process: cultivation process for maximum production of cell mass

밀폐된 배양용기의 온도 30℃, 산도 7.0, 회전수 250rpm, 공기 투여량 1VVM, 용기압력 0.5 ㎏/㎠ 조건으로 24 - 30시간 배양한다.Incubate at a temperature of 30 ° C. in a closed culture vessel, pH 7.0, rotational speed of 250 rpm, air dose of 1 VVM and vessel pressure of 0.5 kg / cm 2 for 24 to 30 hours.

제 4공정 : Formulation 과정Fourth Process: Formulation Process

배양액의 균수를 현미경 및 연속희석 평판 계수법으로 확인 후, 생균수를 4 ∼ 8 ×108cfu/ml로 조정하고 준비된 알긴산과 Glutaraldehyde를 농도 0.2 - 1.0로 배양액에 첨가하여 혼합한다.After confirming the number of bacteria in the culture by the microscope and continuous dilution plate counting method, the number of living cells is adjusted to 4 ~ 8 × 10 8 cfu / ml, and the prepared alginic acid and Glutaraldehyde is added to the culture medium at a concentration of 0.2-1.0 and mixed.

제 5공정 : 포장 및 저장5th process: packing and storage

준비된 용기에 담아 밀폐한 후 저온실에 보관한다.Put it in the prepared container, seal it and store it in the low temperature room.

다음은 본 발명의 안정제 시험결과표와 각종 폐수에 적용한 예를 구체적으로 표시하였다.The following is an example of the stabilizer test result table of the present invention and the examples applied to various wastewaters.

1. 안정제 첨가에 따른 생균수의 변화1. Variation of viable cell number by the addition of stabilizer

경과일수Elapsed days 0.5첨가 시험구0.5 addition test strip (마리/㎖)(Mari / ml) 0.25첨가 시험구0.25 addition test strip (마리/㎖)(Mari / ml) 무처리 대조구Untreated control (마리/㎖)(Mari / ml) 1일1 day 6×108 6 × 10 8 6×108 6 × 10 8 6×108 6 × 10 8 8일8th 7×108 7 × 10 8 3×108 3 × 10 8 2×108 2 × 10 8 15일15th 6×108 6 × 10 8 1×108 1 × 10 8 4×107 4 × 10 7 22일22 days 6×108 6 × 10 8 9×107 9 × 10 7 1×107 1 × 10 7 29일29 days 5×108 5 × 10 8 7×107 7 × 10 7 8×106 8 × 10 6 36일36 days 5×108 5 × 10 8 2×107 2 × 10 7 3×106 3 × 10 6

생균수의 측정은 연속희석에 의한 평판시험법에 의한 결과이다.The number of viable cells is measured by the plate test method by continuous dilution.

2. 방직, 피혁, 식품 및 제지폐수 적용결과표2. Result table for textile, leather, food and paper wastewater

실시번호Conduct number 폐수종류Wastewater Type 초기 CODInitial COD (ppm)(ppm) 10일 경과후 CODCOD after 10 days (ppm)(ppm) 처리효율Processing efficiency ()() 처리구Treatment 무처리구No treatment 처리구Treatment 무처리구No treatment 1One 방직폐수Textile Wastewater 158158 3232 114114 8282 2828 22 피혁폐수Leather wastewater 350350 4646 306306 8787 1313 33 제지폐수Paper wastewater 10801080 176176 970970 8484 1010 44 식품폐수Food wastewater 650650 8484 620620 8787 55

폐수처리 Pilot Test의 운전조건은 실온(18∼25℃)에서 용존 산소농도 1 ∼ 2 ppm, 운전방식은 회분식으로 실시하였다.The wastewater treatment pilot test was run at room temperature (18 ~ 25 ℃) with dissolved oxygen concentration of 1 ~ 2 ppm, and the operation was carried out batchwise.

본 발명으로 얻어진 액상 종균제의 COD제거율은 폐수의 종류에 따라 다소 편차가 있지만 실시한 모든 폐수에 대해 80이상의 높은 처리효율을 나타내었고 무처리구에 비해 월등한 효과를 볼 수 있었다.The COD removal rate of the liquid spawning agent obtained by the present invention was slightly varied depending on the type of wastewater, but showed high treatment efficiency of more than 80 for all wastewaters carried out, and was superior to the untreated section.

Claims (5)

Serratia, B. subtilis 및 광합성 세균을 각각 독립된 배양조에서 최적으로 자랄 수 있도록 배지, 산도 및 온도를 조절하여 충분히 배양한 배양액과 폐수처리 시 스템 내에 부족한 소량의 황산마그네슘, 이인산소다를 첨가하고 균주의 유통 안 정성을 위해 일정 농도의 알긴산과 Glutaraldehyde를 첨가한 액상 종균제의 제조방법.In order to grow Serratia, B. subtilis and photosynthetic bacteria in independent culture tanks, the medium, acidity, and temperature were adjusted to add sufficient amount of magnesium sulfate and sodium diphosphate in the culture solution and wastewater treatment system. Process for preparing liquid spawn with addition of a certain concentration of alginic acid and Glutaraldehyde for distribution stability. 액상 종균제에 사용되는 균주로서Serratia,B. subtilis및 광합성 세균인Rhodopseudomonassp.를 이용한 액상 종균제의 제조방법.A method for preparing a liquid spawn using Serratia , B. subtilis and photosynthetic bacteria Rhodopseudomonas sp. As strains used in the spawn spawn. 균주의 안정성 및 내용물의 보존을 위해 0. 2 - 1.0 범위에서 알긴산을 첨가한 액상 종균제의 제조방법.Method for producing a liquid seed agent added alginic acid in the range of 0.2 to 1.0 for the stability of the strain and the preservation of the contents. 액상 종균제내의 균주들의 성장을 억제하여 유통 안정성을 확보해 주도록 0.2 - 1범위에서 25농도의 Glutaraldehyde를 사용한 제조방법.Method of using 25 concentrations of Glutaraldehyde in the 0.2-1 range to ensure the stability of the distribution by inhibiting the growth of strains in the liquid spawn. 첨가되는 미생물 및 폐수내 존재하는 미생물의 성장촉진을 위해 0.2 - 5범위로 황산 마그네슘과 이인산소다를 첨가하는 액상 종균제의 제조방법Method for preparing a liquid spawn agent which adds magnesium sulfate and sodium diphosphate in the range of 0.2-5 to promote growth of added microorganisms and microorganisms present in wastewater
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100449692B1 (en) * 2002-04-08 2004-09-22 자연과함께 주식회사 Manufacture method of substances for preserving liquid biological products
KR100850373B1 (en) * 2007-01-19 2008-08-04 이화여자대학교 산학협력단 Serratia sp. sy5 and method of plant growth promoting and soil purification using the same
KR100960401B1 (en) * 2009-01-23 2010-05-28 로하스코리아 주식회사 Method for manufacturing water treatment agent used to improve the environment, and water treatment agent made thereby
KR101486120B1 (en) * 2011-09-14 2015-01-26 고려대학교 산학협력단 Method for removing heavy metal using siderophore produced by microbes
CN112300967A (en) * 2020-11-16 2021-02-02 佛山市顺德区活宝源生物科技有限公司 Photosynthetic bacteria pond head expanding culture medium and method for preparing photosynthetic bacteria liquid by using same

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100449692B1 (en) * 2002-04-08 2004-09-22 자연과함께 주식회사 Manufacture method of substances for preserving liquid biological products
KR100850373B1 (en) * 2007-01-19 2008-08-04 이화여자대학교 산학협력단 Serratia sp. sy5 and method of plant growth promoting and soil purification using the same
KR100960401B1 (en) * 2009-01-23 2010-05-28 로하스코리아 주식회사 Method for manufacturing water treatment agent used to improve the environment, and water treatment agent made thereby
KR101486120B1 (en) * 2011-09-14 2015-01-26 고려대학교 산학협력단 Method for removing heavy metal using siderophore produced by microbes
CN112300967A (en) * 2020-11-16 2021-02-02 佛山市顺德区活宝源生物科技有限公司 Photosynthetic bacteria pond head expanding culture medium and method for preparing photosynthetic bacteria liquid by using same

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