LOGO Industrial Microbiology 산업미생물학 Industrial Microbiology 생물환경학과 김정호 Department of Bio-Environmental Science Department of Bio-Environmental Science

 Actinobacteria  Corynebacteria  Lactic acid bacteria  Acetic acid bacteria  Bacillus  Clostridium  Pseudomonas  Escherichia coli  Zymomonas  Archaebacteria 산업적으로 중요한 세균 산업미생물학

Actinobateria ( 방선균 )  Actinomycetes (Ray fungi)  Phylum : Actinobacteria Class : Actinobacteria Order : Actinomycetales  세균과 곰팡이의 중간적 성상 (Filamentous bacteria) 곰팡이 : 균사 형성, 무성포자 형성 균사가 가늠, 미세구조가 세균 세포의 구조와 같음 세균 : 원핵세포, 세포벽은 peptidoglycan  High G+C, Gram (+) bacteria  Soil, fresh water  Antibiotic Actinomycetes : Streptomyces,.. Nitrogen Fixing Actinomycestes : Frankia Decomposer Actinomycetes : cellulose, lignin, chitin, … 산업미생물학

산업적 이용  2 차 대사산물 (Secondary metabolite)  항생물질 Streptomyces grieseus : streptomycin Streptomyces venezuelae : chloramphenicol Streptomyces aureofaciens : tetracycline Streptomyces orientalis : vancomycin  면역조절물질, 염증억제물질, …  각종 효소 생산  난분해성 물질 분해 산업미생물학

 중요한 방선균 genus Streptomyces genus Nocardia genus Actinoplanes genus Actinomadura genus Micromonospora * Thermoactnomyces Thermomonospora  Rare Actinomycetes ( 희귀 방선균 ) Non-streptomycete actinomycetes 산업미생물학

 Corynebacteria  Coryne ← corönë (Greek, knotted rod, club)  Phylum : Actinobacteria Order : Actinomycetales  Genus Cornyebacterum Genus Brevibacterium Genus Arthrobacter  Coryne-form bacteria 산업미생물학

 Gram (+) predominance of mesodiaminopimelic many repetitions of arabinogalactan corynemycolic acid  Rod-shaped (straight or slightly curved)  Pleomorphic : 다형성  Snapping fission : 순간분열  Aerobic / Facultative anaerobic  High G+C content : 51~65% 산업미생물학

산업적 이용  세포벽 투과성 조절이 쉬움  Amino acids : glutamic acid (MSG), lysine, threonine, … Nucleotides Nutritional factors Steroid conversion Cheese aging Enzymes Degradation of hydrocarbons Antitumor agents  Corynebacterium glutamicum, …  Pathogen Corynebacterium diphtheriae 산업미생물학

 젖산균 (Lactic acid bacteria, LAB)  당류를 발효하여 젖산을 대량 생산하는 세균  (G+C) 함량이 낮은 그람 양성의 간균 또는 구균 ; Baciillus 강 예외 ) Bifidobacterium : (G+C) 함양이 높은 그람 양성균 Actinobacteria 강  내산성 (acid tolerant)  Generally no aerobic respiration Catalase (-), Cytochrome (-) 통성혐기성, 미소호기성, 내산소성 혐기성, 절대혐기성  영양요구성이 까다로움 (fastidious nutritional requirement)  Normal microbial flora of human intestine 젖산균 산업미생물학

 Lactobacillus Leuconostoc Pediococcus Lactococcus Streptococcus Bifidobacterium  Aerococcus, Carnobacterium, Enterococcus, Oenococcus, Sporolactobacillus, Tergenococcus, Vagococcus, Weisella 중요한 젖산균 속 산업미생물학

Lactobacillus bulgaricus Streptococcus lactis BifidobacteriumLeuconostoc mesenteroides 산업미생물학

 Under conditions of excess glucose and limited oxygen  Homolactic fermentation ( 동종, 정상 젖산발효 ) C 6 H 12 O 6 (glucose) → 2CH 3 CHOHCOOH (lactic acid)  Heterolactic fermentation ( 이종, 이상 젖산발효 ) C 6 H 12 O 6 → CH 3 CHOHCOOH + C 2 H 5 OH + CO 2  Heterolactic fermentation by Bifidobacterium C 6 H 12 O 6 + 6H 2 O → 2CH 3 CHOHCOOH + 3CH 3 COOH Bifidobacterium : (G+C) 함양이 높은 그람 양성균  Pseudo lactic fermentation by Escherichia coli 2C 6 H 12 O 6 + H 2 O → 2CH 3 CHOHCOOH + C 2 H 5 OH + CH 3 COOH + 2CO 2 + 2H 2 E. coli : Gram (-), pseudo-lactic acid bacteria LAB metabolism 산업미생물학

 Homo-fermentative (Homolactic) LAB  Embden–Meyerhof-Parnas (EMP) pathway C 6 H 12 O 6 (glucose) → 2CH 3 CHOHCOOH (lactic acid)  Streptococcus, Pediococcus, Enterococcus, Lactococcus 속 Group I Lactobacillus (L. delbruekii, L. bulgaricus, L. acidophilus, L. casei, L. plantarum)  Hetero-fermentative (Heterolactic) LAB  Pentose phosphate pathway C 6 H 12 O 6 → CH 3 CHOHCOOH + C 2 H 5 OH + CO 2  Leuconostoc 속 Group III Lactobacillus (L. brevis, L. kefir, L. buchneri, L. fermentum, L. pentaoaceticus) 산업미생물학

 GRAS (Generally Recognized As Safe)  발효식품 제조  김치, 장류, 주류, 요구르트, 치즈, 소시지 등  Leuconostoc kimchii, Leuconostoc mesenterioids, Lactobacillus bulgaricus, Bifidobacterium Leuconostoc oenos (Malo-Lactic fermentation)  젖산 제조  Homolactic LAB  Lactobacillus delbruekii, Streptococcus lactis, Lactobacillus acidophilus  Dextran 생산 Leuconostoc mesenteroides 젖산균의 이용 산업미생물학

 Probiotics  Living organisms which upon ingestion in certain numbers exert health benefits beyond inherent basic nutrition. 5x10 9 CFU/day for at least five days 젖당 소화, 변비, 설사, 면역능력 증강, 콜레스테롤 저하, 암 예방, …  Eli Metchinikoff : Nobel laureate immunologist 1908 : 빌칸 지역 농민의 장수 이유 : 발효 유제품 섭취 Lactobacillus bulgaricus, Streptococcus thermophilus  Lactobicillus acidophilus, Lactobacillus GG, Lactobacillus casei, Streptococcus faecalis, Bifidobacteriu bifidum, …  Prebiotics Synbiotics 산업미생물학

 Bacteriocin  proteinaceous toxins produced by bacteria to inhibit the growth of similar or closely related bacterial strain(s)  Nisin (Lactococcus lactis, 천연식품보존제로 사용 허가 ) Lactacin, Pediocin, …  Streptokinase  혈전용해제  Streptococcus pyogenes 산업미생물학

 식품 부패균 Lactobacillus homohoichii, Lactobacillus heterohoichii, Pediococcus cerevisiae, Lactobacillus thermophilus  Pathogens  Streptococcus pyogenes : 화농성 연쇄상구균  Streptococcus pneumoniae : 폐렴 연쇄상구균  Streptococcus mutants : 치석균 산업미생물학

초산균  초산균 (Acetic acid bacteria, AAB)  당, ethanol 을 산화하여 acetic acid 생성 CH 3 CH 2 OH + O 2 → CH 3 COOH + H 2 O 1. Alcohol dehydrogenase CH 3 CH 2 OH + 1/2 O 2 + NADH 2 → CH 3 CHO + H 2 O + NAD 2. Aldehyde dehydrogenase CH 3 CHO + NADP+ + H 2 O → CH 3 COOH + NADPH 2  Class : α-Proteobacteria, Order : Rhodospiralles, Family : Acetobacteriaceae  Gram (-), Rod-shaped Obligate aerobic, Catalase (+), Oxidase (-)  Optimum pH 3.5 ~ 6.3  주로 식초 (vinegar) 생산에 사용 산업미생물학

 Genus Acetobacter  Acetic acid 생성력 강함  Acetic acid 를 CO 2 와 H 2 O 로 산화 (full set of TCA cycle enzymes)  Genus Gluconobacter  Acetic acid 생성력 약함  Acetic acid 를 CO 2 와 H 2 O 로 산화하지 못함  Polyalcohol 로부터 Keto-acid 생성력 강함 (Glucose → Ketogluconic acid) Acetobacter aceti Gluconobacter oxydans 산업미생물학

 Gluconic acid-producing bacteria ( 글루콘산균 )  Glucose 로부터 다량의 gluconic acid 생성  Gluconobacter spp. Acetobacter xylinum, A. aceti, A. oxydans, …  Sorbose-producing bacteria ( 솔보오스균 )  D-sorbitol 에서 L-sorbose 생산  L-sorbose : vitamin C 생산  Gluconobacter oxydans, Acetobacer aceti, … 산업미생물학

 Genus Bacillus  Class : Bacilli, Order : Bacillales, Family : Bacillaceae  Gram (+), Rod-shaped  Obligate aerobic or facultative anaerobic, Catalase (+)  Endospore-forming  Widely distributed in nature Bacillus subtilis Bacillus megaterium Endospore Bacillus 산업미생물학

 Industrially important  가수분해효소 : amylase, protease, …  항생물질 : polymyxin, bacitracin, gramicidin, subtilin, …  살충제 : BT toxin (Bacillus thuringiensis, …)  제한효소 : BamH1  Model system  분화 (Differentiation) : 포자 형성 / 발아  Bacillus subtilis  Pathogen  Bacillus anthracis : anthrax ( 탄저병 )  Bacillus cereus : 식중독  Bacillus coagulans : 식품 부패 산업미생물학

 Bacillus as a host cell in Genetic engineering  GRAS (Generally Recognized As Safe)  형질전환 가능  단백질을 쉽게 세포 밖으로 분비 : Gram (+)  Plasmid 불안정성  단백질 분해 : protease deficient strains  Paenibacillus  formerly Bacillus (Bacillus polymyxa → Paenibacillus polymyxa)  항생물질 생산  Pattern forming Paenibacillus dendritiformis 산업미생물학

 Genus Clostridium  Class : Clostridia, Order : Clostridiales Family : Clostridiaceae  Gram (+), Rod-shaped Clostridium ← kloster (Greek, spindle)  Obligate anaerobic  Endospore-forming Clostridium botulinum C. perfringens C. tetanus Clostridium 산업미생물학

 Industrially important  Organic solvent : Acetone-butanol fermentation (C. acetobutyricum) Acetic acid, butyric acid, 1,3-propanediol, …  Hydrolytic enzymes : amylase, cellulase, … Clostridium thermocellum : cellulase (bioethanol)  의약품 생산 Clostridium botulinum : Botox  유기염소계 화합물 ( 농약 ) 탈염소화 (dechlorination)  Pathogen  Clostridium botulinum : 통조림 멸균시 기준  Clostridium perfringens : 식중독  Clostridium tetani : tetanus ( 파상풍 ) 산업미생물학

 Genus Pseudomonas  Class : Gamma Proteobacteria, Order : Pseudomonadales, Family : Pseudomonadaceae  Gram (-), Rod-shaped  One or more polar flagella : motile  Aerobic : catalase (+)  Non-spore forming  Some are psychrophilic : 냉장식품의 부패 (protease, lipase) Pseudomonas 산업미생물학

 특징  다양한 서식 환경  다양한 대사경로  동 ∙ 식물 병원균 많음 Pseudomonas aeruginosa ( 녹농균 ) 항생제 저항성  수용성 형광색소 생산균 있음 Pseudomonas fluorescence 산업미생물학

 산업적 이용  PHAs (Poly-β-hydroxyalkanoates) 생분해성 플라스틱 Pseudomonas oleovorans  Bioremediation agents 난분해성 물질의 분해 P. alcaligenes : Polycyclic aromatic hydrocarbons  Biocontrol agents Pseudomonas fluorescence  빙핵활성 단백질 (ice nucleation protein) Pseudomonas syringae 산업미생물학

고세균 (Archaea)  Archaea (singular archaeon)  a domain (kingdom) of single-celled microorganisms  Prokaryotic : no nucleus, no membrane-bound organelles Visual similarity to bacteria Asexual reproduction by binary fission (or fragmentation, budding)  Ether lipids in cell membranes (archaeol) No peptidoglycan in cell wall : pseudopeptidoglycan rRNA different from eubacteria Transcription and translation – similar to eukaryotes  Many are extremophiles  No known species forms spores  No clear examples of archaeal pathogens or parasites 산업미생물학

고세균의 이용  Extremophile  resistant to heat, acidity, alkalinity, salt enzymes that function under harsh conditions  Thermostable enzymes Pfu DNA polymerase (Pyrococcus furiosus, PCR) amylases, galactosidases, pullulanases very stable in organic solvents (green chemistry)  Sewage treatment & biogas production methanogenic archaea  Extraction of metals (gold, cobalt, copper) acidophilic archaea  Antibiotics (archaeocins) 산업미생물학