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EN27435 ENVIRONMENTAL MICROBIOLOGY Fall 2005 Prof. Changwon Kim

EN27435 ENVIRONMENTAL MICROBIOLOGY Fall 2005 Prof. Changwon Kim 재료관 502 051-510-2416 cwkim@pusan.ac.kr Office Hours: By appointment. Texts Microbiology by Prescott, Harley & Klein 환경미생물학 , 김창원 등 , 동화기술 환경미생물학 , 이상준 등 , 형설 PLEASE BE ON TIME!

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EN27435 ENVIRONMENTAL MICROBIOLOGY Fall 2005 Prof. Changwon Kim

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  1. EN27435 ENVIRONMENTAL MICROBIOLOGY Fall 2005Prof. Changwon Kim 재료관 502 051-510-2416 cwkim@pusan.ac.kr Office Hours: By appointment

  2. TextsMicrobiology by Prescott, Harley & Klein 환경미생물학, 김창원 등, 동화기술 환경미생물학, 이상준 등, 형설 PLEASE BE ON TIME! Grades: based on average of two exams (80%), at least 5 assignments (10%), and attendance (10%) Academic Integrity Violations will not be tolerated. The penalty may be severe. It is your duty to conduct yourself in an appropriate manner.

  3. BIO 3595 GENERAL MICROBIOLOGY Fall 2004Dr. John Friede Mendel 143 9-4833 Office Hours: By appointment

  4. 환경생물공학의 특징 - 혼합균주 - 비살균계 - 성공비결 : ①경쟁생존 ②유효기능유지 ③불필요 미생물 증식 제어 • Environmental Microbiology? Biotechnology • - 정의 • - 적용분야 • Science: 자연현상 원리 발견(물질, 성질, 변환/반응) • Engineering: 발견된 자연현상의 원리 등을 인위적으로 활용 • (반응속도, 에너지 보존법칙)

  5. MICROBIOLOGY - the study of little things a branch of biology VIROLOGY - viruses BACTERIOLOGY - bacteria MYCOLOGY - fungi and yeast PROTOZOOLOGY - protozoa PARASITOLOGY IMMUNOLOGY

  6. MAJOR DEVELOPMENTS Development of the Microscope Antonie van Leeuwenhoek - 1676 Improved lenses – late 1800s Electron microscope

  7. MAJOR DEVELOPMENTS Dispelling of spontaneous generation maggots on meat; spoiling of broth; rats Francesco Redi – mid 1600s cover meat in jars Louis Pasteur – mid 1800s boil broth in gooseneck flask role of microbes in fermentation Acceptance of role of microbes in disease Adoption of Aseptic Technique Vaccination & Immunology Discovery of sulfa drugs & penicillin

  8. Genetic Engineering - The newest technology! Clean vaccines egg allergies New antibiotics

  9. Cell Structure & Function

  10. CLASSIFICATION - 5 Kingdoms (Whittaker, 1969) Prokaryotae or Monera Protista Fungi Plantae Animaliae

  11. CLASSIFICATION - 5 Kingdoms (Whittaker, 1969) Prokaryotae or Monera Protista Fungi Plantae Animaliae CLASSIFICATION - 3 Kingdoms (Woese, 1978) Eubacteria - true bacteria Archaebacteria - ancient bacteria Eukaryotes - protists, fungi, plants, animals

  12. BASICS OF CLASSIFICATION Morphological Characteristics cell structure Staining Properties Biochemical Characteristics Serological Properties Genetic Characteristics Proteins

  13. NOMENCLATURE Binomial nomenclature Genus and species; e.g. Homo sapiens Classification Heirarchy Kingdom Phylum Class Order Family Genus Species

  14. 복잡화학계(complex chemical system) ① 성장, 번식 : 독립체 생성 ② 주위 환경보다 낮은 엔트로피 상태; 고도록 조직화 ③ 화학적 환원원소(C, N, P, S, H, O)로 구성; 유기물 ④ 원소, 전자, 에너지를 얻는 대사작용 Cell 세 포 • 정의: 생명체 기본단위, • 독립체, • 복잡한 화학적 계(system)

  15. prokaryotes(원핵생물) : 단세포, 핵막 및 핵 속에 염색체 없음 • pro = before karyon = nucleus bacteria(박테리아) archaea(고세균) • eukarya(진핵생물) : 단세포 혹은 다세포, 핵속에 염색체 있음, • 고등생물 • eu = true karyon = nucleus algae(조류) protista(원생생물) : • 환경생물공학의 관심 영역: 박테리아, 고세균 • 진핵생물 중 조류, 원생동물 • 최근 식물 단세포 미생물

  16. 원핵세포와 진핵세포의 구조 Click here : Go Back 1.1

  17. 세포 구성요소 • 세포벽(cell wall) : 막 보호, 세포 견고성 확보 • 세포막(plasmamembrane) : 주위환경과 경계, 물질전달 경로 • 세포질(cytoplasm) : 물+기능성 거대분자, 세포내부구성 • 핵양체(nucleoid) : 유전물질(DNA) 보관, 영양물질 가공 • 염색체(chromosome) : 유전적 정보 보유(DNA) • 플라스미드(Plasmid) : 염색체와 따로 증식할 수 있는 DNA • - 리보솜(ribosome) : 단백질 합성 (RNA+protein)

  18. 분류학(taxonomy)과 계통발생론(phenotype)에 의한 미생물 분류

  19. PROKAYOTIC CELL 원핵세포 • 박테리아 (bacteria) : 유기물 → 아세트산, 수소 • 고세균 (archaebacteria) : 유기물 → 아세트산, 수소 → 메탄 • 방선균 (actinomycetes) : 곰팡이 처럼 균사 형성 • 남조류 (bluegreen algae, cyanobacteria) : 맛, 냄새, 독소

  20. PROKARYOTIC CELL Shapes: rod (bacillus, bacilli) sphere (coccus, cocci) spiral (spirillum, spirilli) pleomorphic Cell arrangement: single cell pairs (diplococcus) chains clusters

  21. PROKARYOTIC CELL STRUCTURE Capsule or slime layer protein or carbohydrate non-essential under genetic control under environmental controlS. salivariusE. aerogenesS. pneumoniae function protection: water; phagocytes attachment: surfaces; other cells

  22. PROKARYOTIC CELL STRUCTURE Fimbria (fimbriae) &: Pilus (pili) fimbriae shorter, more numerous tubular structures fimbria – attachment other bacterial cells - pellicle other cells – gonococcus evade phagocytes pilus – conjugation plasmid transfer

  23. PROKARYOTIC CELL STRUCTURE Flagellum – locomotion location and number vary monotrichous polytrichous peritrichous structure filament hook basal granule

  24. PROKARYOTIC CELL STRUCTURE Flagellum – locomotion filament – single protein, flagellin H antigen protein subunits globular tubular structure helical shape 2.3 µm pitch, left-handed helix hook – anchor filament to basal granule chemically different basal granule – central rod with 2-4 rings Gram-positive 2 rings Gram-negative 4 rings M ring – membrane associated S ring – just above membrane; anchored to wall P ring – associated with peptidoglycan L ring – associated with outer layer of envelope

  25. PROKARYOTIC CELL STRUCTURE Flagellum – locomotion basal granule L ring P ring S ring M ring

  26. PROKARYOTIC CELL STRUCTURE Flagellum – locomotion Motion – rotation of filament by basal granule (M ring) motion types: swimming, tumbling swimming – counterclockwise rotation tumbling – clockwise rotation Energy source – membrane potential ΔµH+ H+ flow through basal granule M ring rotates swimming vs. tumbling methylation of key component

  27. PROKARYOTIC CELL STRUCTURE Cell wall – mucopeptide, murein, peptidoglycan external layer, responsible for shape rod (bacillus), sphere (coccus), spiral (spirillum) Archaebacteria - different Eubacteria – two types, unlike plant wall staining properties different with Gram stain

  28. PROKARYOTIC CELL STRUCTURE Cell wall – mucopeptide, murein, peptidoglycan composition Gram-positive Gram-negative teichoic acid 10-40% none polysaccharide <10% 10-15% lipid none 20% mucopeptide 50-90% 15% protein none 50%

  29. Teichoic acids polyol – phosphate backbone polyols: glycerol or ribitol other substituents: sugars, amino sugars, amino acids backbone linked to mucopeptide via muramic acid function – sequester divalent cations evidence --

  30. Teichoic acids - Ribitol P O H-C-H H-C-OH D-Ala H-C-OH H-C-OH (glucose, glucosamine, amino acids) H-C-H O P

  31. Lipopolysaccharide – LPS only in Gram-negative organisms in outer layer of envelope (outer membrane) antigen – O antigen “ohne hauch” pyrogenic endotoxin - fever, edema, diarrhea, shock, internal hemorrhage structure: lipid A, R core, O antigen lipid A: fatty acids core: KDO, heptoses O antigen: variable, odd sugars (3-deoxy, 6-deoxy, 3,6-dideoxy) rough mutants function: outer membrane integrity permeability, protect against phagocytes

  32. Protein lipoprotein (Braun’s lipoprotein) structural component of envelope porin proteins pores in outer membrane three subunits selective permeability

  33. Mucopeptide structure & composition varies between organisms (G+ vs G-) composition: N-acetylmuramic acid N-acetylglucosamine amino acids: alanine glutamic acid lysine or diaminopimelic acid

  34. Mucopeptide structure & composition varies between organisms (G+ vs G-) composition: N-acetylmuramic acid (1) N-acetylglucosamine (1) amino acids: alanine (2) glutamic acid (1) lysine or diaminopimelic acid (1)

  35. Mucopeptide structure & composition varies between organisms (G+ vs G-) composition: N-acetylmuramic acid (1) N-acetylglucosamine (1) amino acids: alanine (2) 1 D-Ala and 1 L-Ala glutamic acid (1) D-Glu lysine or diaminopimelic acid (1) L-Lys Penicillin and wall synthesis Park nucleotide 3 alanines – 2 D-Ala and 1 L-Ala

  36. Mucopeptide Precursor – UDP carrier Park nucleotide intermediate

  37. Mucopeptide subunit structure

  38. Mucopeptide

  39. Mucopeptide – most Gram-negative, many Gram-positive Fig. 23.4

  40. Mucopeptide – many Gram-positive (Fig. 23.4) 1. S. aureus 4. Lactobacillus viridescens2. S. epidermidis 5. Streptococcus salivarius3. Micrococcus roseus 6. Leuconostoc cremoris

  41. Mucopeptide – unusual variation

  42. 그람염색 (Gram Stain)

  43. Gram-positive wall

  44. Gram-negative cell envelope

  45. Cytoplasmic membrane Phospholipid bilayer with proteins Lipids – eubacteria: ester-linked lipids, no sterols archaebacteria: ether-linked lipids, sterols

  46. Membrane function - permeability electron transport chain transport diffusion – no carrier, no energy equilibrium: [in] = [out] facilitated diffusion – carrier, no energy equilibrium: [in] = [out] transport – carrier, energy equilibrium: [in] >> [out]

  47. Transport systems common characteristics stereospecific – single compound or closely related family leucine, isoleucine, valine concentrate against a gradient gradients of 600:1 to 2000:1 [in] to [out] follow saturation kinetics increase concentration, rate increases to saturation of carriers; higher concentrations, no rate increase

  48. Transport systems energy sources – phosphotransferase system group translocation reaction components: HPr, Enzyme I and Enzyme II HPr & Enz I: cytoplasmic, constitutive Enz II: membrane-associated, inducible reaction: PEP + HPr → pyruvate + P-HPr P-HPr + sugar → HPr + sugar-P phosphorylation and transport are synonymous

  49. Transport systems energy sources – membrane potential ion gradients, exterior positive H+ or Na+ co-transport carrier accepts S only after H+ attaches symport or cotransport outside: C + H+→ C* C* + S → C*S inside: C*S → H+ + CS → C + S

  50. Transport systems energy sources – traffic ATPase binding protein – specific transport protein – common carrier ATP binding & hydrolysis conformational change

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