Lactic Acid
Lactic Acid
Lactic Acid
Mikroorganisme mengeluarkan enzim ekstra seluler untuk memecah karbohidrat Monomer dari karbohidrat bisa langsung masuk kedalam sel Ada 3 jalur pemecahan glukosa menjadi asam piruvat Bakteri asam laktat homofermentatif digunakan dalam pengawetan makanan
Jelaskan
Kelompok bakteri asam laktat Homofermentatif Heterofermentatif
Polyhydroxyalkanoates (PHAs)
Lactobacillus
Gram + Short Rods Homo or hetero fermentative All ferment frucose Most produce lactic acid from D/L Glucose Produce sour taste
Oenococcus
Gram + Heterofermentative Can be difficult to distinguish from Lactobacillus under a microscope Produce lactic acid from malic acid Makes mannitol from fructose Makes lactic acid from glucose Diacetyl character is most balanced compared to other lactics
Pediococcus
Gram + Form spherical cells in tetrads Aerobic to microaerophillic Homofermentative Can produce acetate, diacetyl Complex nutrient requirements Can produce ropiness sensory character but is rare
Bacteria Basics
Homofermentative vs. Heterofermentative Gram + vs. Gram Oxygen usage
Antiport
Sugar1 Sugar2
PEP-PTS
Sugar
Sugar
F1F0 ATPase
Permease
Permease
EIIBCS
EIIAS
IN
ATP ADP
+ H
Sugar Sugar1 Sugar2
HPr
+ H
EI
Sugar-PO4
PEP Energy from: PMF (pH + ) Concentration Gradient (S1 & S2)
Pyruvate
PEP
PEP-PTS
OUT
Glucose
Galactose
Lactose Permease
Glucose Permease
Galactose Permease
IN H
+
Lactose
+
Glucose
+
Galactose
Galactose
Lactose H+
H+ Lactose
Glucose
Glucose-6-phosphate Fructose-6-phosphate
ATP ADP
Fructose-1,6-diphosphate
ATP ADP
FDP aldolase
Dihydroxyacetone-phosphate (2) Glyceraldehyde-3-phosphate (2) H2PO4(2) 1,3-Diphosphoglycerate (2) NAD+ (2) NADH + (2) H+
(2) 3-Phosphoglycerate
Products: 2 ATP 2 Lactate Key enzymes: FDP aldolase Lactate dehydrogenase (2) 2-Phosphoglycerate
(2) H2O (2) Phosphoenolpyruvate (PEP) (2) Pyruvate (2) ADP (2) ATP (2) NADH + (2) H+ (2) NAD+
(2) Lactate
cofactor regeneration
FDP aldolase
Dihydroxyacetone-phosphate (2) Glyceraldehyde-3-phosphate (2) H2PO4(2) NAD+ (2) NADH + (2) H+ (2) ADP (2) ATP (2) ADP (2) ATP
Homolactic
(LDH)
(2) Lactate
(2) NAD+
(2) NADH + (2) H+ H2PO4Products: 3 ATP 2 Formate 1 Ethanol 1 Acetate Key enzymes: FDP aldolase Pyruvate formate lyase (PFL)
(2) Pyruvate
(2) CoA
(2) Formate
CoA NADH + H+ Acetaldehyde NAD+ NADH + H+ NAD+
Acetate
Ethanol
cofactor regeneration
Homolactic fermentation prevails when glucose is abundant. Mixed acid fermentation prevails when sugars are limited semi-starvation.
Mixed acid fermentation prevails during growth on galactose as the primary sugar source.
Homolactic fermentation prevails under aerobic conditions because the pyruvate formate lyase (PFL) enzyme is oxygen sensitive.
Glucose
Glucose-6-phosphate 6-phospho-gluconate
NADH +
H+
O2
NADH oxidase
H2O2
NAD+
NADH + H+ NAD+
CO2
NADH oxidase
2 H2 O
Phosphoketolase
Glyceraldehyde-3-phosphate H2PO4
-
Acetate
Products: 2 ATP 1 CO2 1 Lactate 1 Acetate Key enzymes: Phosphoketolase Lactate dehydrogenase NADH oxidase
LDH
Lactate
Glucose
Glucose-6-phosphate 6-phospho-gluconate
CO2
Phosphoketolase
Glyceraldehyde-3-phosphate H2PO41,3-Diphosphoglycerate 3-Phosphoglycerate 2-Phosphoglycerate
Acetyl-phosphate NAD+ NADH + H+ ADP ATP CoA Acetyl-CoA CoA NADH + H+ NAD+ H2PO4-
Acetaldehyde
(2) H2O
NADH + H+ NAD+
Ethanol
Products: 1 ATP 1 CO2 1 Lactate 1 Ethanol Key enzymes: Phosphoketolase Lactate dehydrogenase
LDH
Lactate
Glucose-6-phosphate
Fructose Fructose-6-phosphate
Glucose-6-phosphate
6-phospho-gluconate
Leloir Pathway
PEP Pyruvate EI EI
Tagatose Pathway
Galactose-6-PO4 Tagatose-6-PO4 Tagatose-1,6-diPO4
ATP ADP
Dihydroxyacetonephosphate
(2) Glyceraldehyde3-phosphate
* Mixed acid fermentation dominates if galactose is the most abundant sugar available
lactose
sucrose
maltose
glucose
fructose
glucose
glucose
galactose
glucose
-galactosidase
Leloir pathway Homolactic, mixed acid, or heterolactic pathway
PO4
PO4
+ Phospho--galactosidase
Tagatose pathway
Permease
IN Glucose
Homolactic, mixed acid or heterolactic pathway
Lactose Galactose
-galactosidase
* S. thermophilus, and Lb. delbrueckii do not metabolize the galactose part of lactose. They export galactose from the cell.
(Figure from Fox et al. 1990. Critical Reviews in Food Science and Nutrition. 29:237-253.)
Tagatose Pathway
Homolactic Pathway
Leloir Pathway
Heterolactic Pathway
Pentoses
Pentose
Pentose Pentose-PO4
isomerization
ATP ADP
Xylulose-PO4 or Ribulose-PO4
Glucose
Glucose-6-phosphate 6-phospho-gluconate
CO2
Ribulose-5-phosphate Xylulose-5-phosphate H2PO4-
O2
NADH oxidase
H2O2
NADH oxidase
2 H2 O
Phosphoketolase
Glyceraldehyde-3-phosphate H2PO4
-
Acetate
Products: 2 ATP 1 Lactate 1 Acetate
LDH
Lactate
Obligately homofermentative
Group I Lactobacillus species and a few other species Hexoses are fermented by homolactic fermentation pathway (glycolysis + LDH)
FDP aldolase
Dihydroxyacetone-phosphate (2) Glyceraldehyde-3-phosphate (2) H2PO4(2) 1,3-Diphosphoglycerate (2) NAD+ (2) NADH + (2) H+
(2) 3-Phosphoglycerate
Products: 2 ATP 2 Lactate Key enzymes: FDP aldolase Lactate dehydrogenase (2) 2-Phosphoglycerate
(2) H2O (2) Phosphoenolpyruvate (PEP) (2) Pyruvate (2) ADP (2) ATP (2) NADH + (2) H+ (2) NAD+
(2) Lactate
cofactor regeneration
Glucose
Glucose-6-phosphate 6-phospho-gluconate
NADH +
H+
O2
NADH oxidase
H2O2
NAD+
NADH + H+ NAD+
CO2
NADH oxidase
2 H2 O
Phosphoketolase
Glyceraldehyde-3-phosphate H2PO4
-
Acetate
Products: 2 ATP 1 CO2 1 Lactate 1 Acetate Key enzymes: Phosphoketolase Lactate dehydrogenase NADH oxidase
LDH
Lactate
Obligately heterofermentative
Group III Lactobacillus species, Leuconostoc, Oenococcus
Facultatively heterofermentative
Group II Lactobacillus species, Lactococcus, Pediococcus, Streptococcus thermophilus
Lactobacillus
Lb. rhamnosus
Homolactic and facultatively heterolactic no CO2 from glucose, FDP aldolase present Obligately heterolactic CO2 from glucose, phosphoketolase present
Citrate Transport
H+ Citrate
Citrate Permease
H+ Citrate
CH2 - COOH
HO - C - COOH CH2 - COOH
Citrate Metabolism
CH2 - COOH
HO - C - COOH
CH2 - COOH
citrate lyase
Acetate CO2
citrate
O O CH3 C C CH3
NADH + H+ NAD+
Leuc. mesenteroides subsp. cremoris Lc. lactis subsp. lactis biovar. diacetylactis
Synthesis of polysaccharides
Fermentasi karbohidrat
Polisakarida dipecah menjadi gula sederhana sebelum difermentasi Tahap pertama fermentasi, glukosa diubah menjadi asam piruvat Tahap kedua fermentasi, asam piruvat diubah menjadi produk akhir yang lebih spesifik
Jalur Heksosamonofosfat (HMP) menghasilkan pentosa yang diperlukan untuk sisntesis asam nukleat, beberapa asam amino aromatik dan vitamin, dilakukan beberapa organisme Glukosa + 12 NADP+ + ATP 6CO2 + 12 NADPH + 12 H+ + ADP + Pi Jalur Fosfoketolase (FK) ditemukan pada bakteri yang tergolong laktobasili heterofermentatif Glukosa + NAD+ + ADP + Pi Piruvat + etanol + CO2 + NADH + H+ + ATP