细菌抗菌肽的分类作用机理及基因重点工程专项研究进展.docx
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细菌抗菌肽的分类作用机理及基因重点工程专项研究进展.docx
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细菌抗菌肽的分类作用机理及基因重点工程专项研究进展
细菌抗菌肽分类、作用机理及基因工程研究进展
刘小雨
10月
当前,由于抗生素在临床治疗中不规范使用,不但导致经济上巨大损失和药物不良反映;并且导致细菌和真菌等病原微生物正在以惊人速度对抗生素产生强耐药性。
特别是多重耐药菌和超级耐药菌浮现,经常在很短时间内浮现全世界范畴内流行,给人们生命安全和经济带来巨大损失。
因而对具备高抗菌活性和安全性抗菌肽研发日益受到注重,人们盼望抗菌肽成为抗生素抱负代替品之一。
抗菌肽(AMPS)普通由10~60个氨基酸残基构成,分子量约为~7000Da,并且热稳定性较好。
抗菌肽抗菌机制不同于老式抗生素,其作用靶点大体分为细胞膜作用靶点和胞内作用靶点(DNA,RNA,酶等)两大类,抗菌肽抗菌活性和毒性与抗菌肽二级构造、净电荷数量和疏水性等理化参数密切有关。
迄今为止,大概有2700各种抗菌肽已经被分离出来,某些已经通过化学合成和基因工程表达大规模制备。
在实际应用方面,某些抗菌肽已成功应用于医疗卫生和食品加工等行业,其中特别引人注意是各种抗菌肽已经进入二、三期临床实验阶段【~】。
抗菌肽来源与制备大体分为三条途径:
天然产物(微生物,植物,动物,海洋生物来源)提取,化学合成(液相合成法和固相合成法)和基因工程(大肠杆菌,枯草芽孢杆菌和毕赤酵母表达载体等)制备。
天然产物提取易受来源限制,特别是来自珍稀动植物资源抗菌肽,提取更易受到原材料来源限制。
化学合成生产成本较高,且有各种剧毒性试剂(氢氟酸、氰化钾等)需要大量应用,产生“三废”对环境导致污染问题也难以解决。
通过基因工程和发酵工程办法制备抗菌肽可以有效减少生产成本,生产工艺容易放大;产生废液解决工艺较为成熟。
当前使用原核表达系统制备抗菌肽占所有抗菌肽基因表达系统80%左右。
人们使用原核表达系统表达抗菌肽已经积累了许多经验,对抗菌肽表达质粒构建,生产工艺中试和放大也有所进展。
但是抗菌肽原核表达依然具备很大技术挑战性,其不但对宿主菌有毒性,并且由于抗菌肽分子量较少,易被宿主菌各种内源性蛋白酶降解。
以往研究表白抗菌肽采用与融合标签表达是实现抗菌肽高效表达有效方略之一。
当前惯用融合标签有硫氧还蛋白(Trx)、类固醇异构酶(KSI)及谷胱甘肽-S-转移酶(GST)等,但在实际应用中,发现上述标签存在标签蛋白过大、促可溶性效果差、表达产物剪切效率低等问题,因而寻找提高抗菌肽可溶性表达和产率办法依然是当前研究重要方向。
1抗菌肽分类
按照抗菌肽来源分类,截止到7月,共有2717种抗菌肽被发布,其中来源于细菌有269种,来源于古生菌有4种,来源于原生生物有8种,来源于真菌有13种,来源于植物有333种,来源于动物有2042种。
由于在芽孢杆菌表达系统中,缺少真核以及动物、植物等相应修饰系统,来源于动物、植物以及真菌基因在原核表达系统中很难得到高效表达,因此本综述中重点关注来自于细菌抗菌肽及其抗菌活性;其详细信息如下:
1.1来源于细菌抗菌肽
来源于细菌抗菌肽一共有269种,其某些信息如下:
Name
Source
Activity
Name
Source
Activity
Hominicin
Staphylococcushominis MBBL2-9
anti-Gram+
anti-Gram-
TricyclicpeptideRP71955
Streptomycesstrains AA3891
Antiviral
anti-HIV
PlantaricinA
Lactobacillusplantarum C11
anti-Gram+anti-Gram-
Cancercells
CarnobacteriocinB2
Carnobacteriumpiscicola
anti-Gram+anti-Gram-
LeucocinA
Leuconostocgelidum UAL-187
anti-Gram+
anti-Gram-
NisinZ
LactococcuslactisSIK-83
anti-Gram+Cancercells
NisinA
Streptococcuslactis
anti-Gram+
Spermicidal
Antibiofilm
Woundhealing,Cancercells
Subtilin
Bacillussubtilis
anti-Gram+
Pep5
Staphylococcusepidermidis strain5
anti-Gram+
MicrocinJ25
Escherichiacoli AY25
anti-Gram-
GramicidinA
soilbacterium,
Bacillusbrevis
(短杆菌)
anti-Gram+anti-Gram-
antiviral
Spermicidal
anti-HIV
EricinS
Bacillussubtilis A1/3
anti-Gram+
SiamycinII
Streptomycesstrains AA3891(链霉菌)
Antiviral
anti-HIV
Lactococcin972
Lactococcuslactis IPLA972(乳酸乳球菌)
anti-Gram+
MundticinATO6
Enterococcusmundtii ATO6
(蒙氏肠球菌)
anti-Gram+
Piscicolin126/PiscicocinVia
Carnobacteriumpiscicola JG126
(栖鱼肉杆菌)
anti-Gram+
SakacinP
Lactobacillussake Lb674
anti-Gram+
PediocinPA-1
Pediococcusacidilactici PAC-1.0
anti-Gram+
Spermicidal
MesentericinY105
Leuconostocmesenteroides
anti-Gram+
CarnobacteriocinBM1
Carnobacteriumpiscicola LV17B
anti-Gram+
anti-Gram-
CurvacinA/sakacinA
Lactobacillussake Lb706
anti-Gram+
Planosporicin
Planomonosporaalba
anti-Gram+
anti-Gram-
GassericinA
Lactobacillusgasseri
Lactobacillusreuteri LA6
anti-Gram+
anti-Gram-
CircularinA
Clostridiumbeijerinckii ATCC25752
anti-Gram+
anti-Gram-
Closticin574
Clostridiumtyrobutyricum ADRIAT932
anti-Gram+
anti-Gram-
EnterocinE-760
Enterococcusdurans/faecium/hirae
anti-Gram+
anti-Gram-
EnterocinA
Enterococcusfaecium
anti-Gram+
DivercinV41
Carnobacteriumdivergens V41
anti-Gram+
DivergicinM35
Carnobacteriumdivergens M35
anti-Gram+
Coagulin
Bacilluscoagulans I-4
anti-Gram+
Listeriocin743A
Listeriainnocua 743
anti-Gram+
MundticinKS
Enterococcusmundtii NFRI7397
anti-Gram+
antiviral
LeucocinC
Lactococcuslactis MMFI
anti-Gram+
LactococcinMMFII
Lactococcuslactis MMFI
anti-Gram+
SakacinG
--
anti-Gram+
Plantaricin423
Lactobacillusplantarum 423
anti-Gram+
PlantaricinC19
Lactobacillusplantarum C19
anti-Gram+
EnterocinP
Enterococcusfaecium P13
anti-Gram+
Bacteriocin31
Enterococcusfaecalis
anti-Gram+
CarnobacteriocinB1
CarnobacteriumpiscicolaLV17B
anti-Gram+
anti-Gram-
ButyrivibriocinAR10
ButyrivibriumfibrisolvensAR10
anti-Gram+
anti-Gram-
SubtilosinA
Bacillussubtilis
Bacillusamyloliquefaciens,or Bacillusatrophaeus
anti-Gram+
anti-Gram
antiviral
Spermicidal
EnterocinAS-48
EnterococcusfaecalisS-48
anti-Gram+
anti-Gram-
PlantaricinASM1
Lactobacillusplantarum A-1
anti-Gram+
Uberolysin
Streptococcusuberis42
anti-Gram+
anti-Gram-
AcidocinB
LactobacillusacidophilusM46
anti-Gram+
anti-Gram-
CarnocinUI49
Carnobacteriumpiscicola UI49
anti-Gram+
anti-Gram-
Lichenin
Bacilluslicheniformis
anti-Gram+
anti-Gram-
NisinQ
HalobacteriumstrainAS7092
anti-Gram+
Serracin-P23kDasubunit
Serratiaplymuthica
anti-Gram+
anti-Gram-
Serracin-P43kDasubunit
Serratiaplymuthica
anti-Gram+
anti-Gram-
Variacin
Kocuriavarians
anti-Gram+
anti-Gram-
MutacinII
Streptococcusmutans
anti-Gram+
anti-Gram-
MutacinIII
Streptococcusmutans
anti-Gram+
Mutacin,orMutacinB-Ny266
Streptococcusmutans
anti-Gram+
anti-Gram-
ThuricinS
Bacillusthuringiensisserovarentomocidus
anti-Gram+
AureocinA53
Staphylococcusaureus A53
anti-Gram+
SiamycinI
StreptomycesstrainAA6532
Antiviral
anti-HIV
NP-06
StreptomycesstrainAA6532
Antiviral
anti-HIV
LactococcinG-a
LactococcuslactisLMGT2081
anti-Gram+
anti-Gram-
LactococcinQ
LactococcuslactisQU4
anti-Gram+
LCI
Bacillussubtilis
anti-Gram+
anti-Gram-
Enterocin1071
EnterococcusfaecalisBFE1071
anti-Gram+
anti-Gram-
PlantaricinS
LactobacillusplantarumLPCO10
anti-Gram+
anti-Gram-
AFP1
StreptomycestendaeTu901(唐德链霉菌)
antifungal
PlantaricinNC8
Lactobacillusplantarum NC8
anti-Gram+
CarnocyclinA
CarnobacteriummaltaromaticumUAL307
anti-Gram+
LactacinF
LactobacillusjohnsoniiVPI11088(Laf+)
anti-Gram+
BrochocinC
BrochothrixcampestrisATCC43754
anti-Gram+
Thermophilin13
Streptococcusthermophilus
anti-Gram+
ABP-118
Lactobacillussalivariussubsp.salivariusUCC118
anti-Gram+
anti-Gram-
SalivaricinP
LactobacillussalivariusDPC6005
anti-Gram+
anti-Gram-
MutacinIV
StreptococcusmutansUA140
anti-Gram+
anti-Gram-
Lactocin705
LactobacilluscaseiCRL705
anti-Gram+
anti-Gram-
Cytolysin
Enterococcusfaecali
(粪肠球菌)
anti-Gram+
anti-Gram-
Mammaliancells
PlantaricinEF
Lactobacillusplantarum C1
anti-Gram+
anti-Gram-
PlantaricinJK
LactobacillusplantarumC1
anti-Gram+
anti-Gram-
EnterocinSE-K4
Enterococcusfaecium
anti-Gram+
AcidocinJ1132
LactobacillusacidophilusJCM1132
anti-Gram+
anti-Gram-
CurvaticinL442
LactobacilluscurvatusL442
anti-Gram+
Bacteriocin32
Enterococcusfaecium
anti-Gram+
Bacteriocin43
Enterococcusfaecium
anti-Gram+
BacteriocinT8
EnterococcusfaeciumT8
anti-Gram+
EnterocinB
Enterococcusfaecium T136; Enterococcusfaecium W3
anti-Gram+
AcidocinA
LactobacillusacidophilusTK9201
anti-Gram+
EnterocinQ
EnterococcusfaeciumL50
anti-Gram+
EnterocinRJ-11
EnterococcusfaecalisRJ-11
anti-Gram+
EnterocinL50
EnterococcusfaeciumL50
anti-Gram+
MR10A
Enterococcusfaecalis MRR10-3; Enterococcusfaecalis 710C
anti-Gram+
anti-Gram-
Lacticin3147
LactococcuslactisDPC3147
anti-Gram+
Spermicidal
SalivaricinA
Streptococcussalivarius20P3
anti-Gram+
MicrocinE492
Klebsiellapneumoniae RYC492
anti-Gram-
Cancercells
HiracinJM79
EnterococcushiraeDCH5
anti-Gram+
Thermophilin9
StreptococcusthermophilusLMD-9
anti-Gram+
PenocinA
PediococcuspentosaceusATCC25745
anti-Gram+
SalivaricinB
StreptococcussalivariusK12
anti-Gram+
Lacticin481
LactococcuslactisCNRZ481
anti-Gram+
BacteriocinJ46
LactococcuslactisJ46
anti-Gram+
NukacinISK-1
StaphylococcuswarneriISK-1,orStaphylococcussimulans3299
anti-Gram+
StreptococcinA-FF22
streptococcusstrainFF22
anti-Gram+
LactocinS
LactobacillussakeL45
anti-Gram+
Mersacidin
Bacillussp.strainHILY-8554728
anti-Gram+
GramicidinB
Bacillusbrevis
anti-Gram+
anti-Gram-
MicrocinC7
Escherichiacoli
anti-Gram-
MicrocinB17
Escherichiacoli
anti-Gram-
MicrocinL
EscherichiacoliLR05
anti-Gram-
MicrocinM
EscherichiacoliMC4100
anti-Gram-
MicrocinI47
EscherichiacoliH47
anti-Gram-
Pep27
Streptococcuspneumoniae
(肺炎双球菌)
anti-Gram+
anti-Gram-
Cancercells
Microcin24
E.coli2424
anti-Gram-
BTL
B.subtilisB-TL2
antifungal
AdDLP
Anaeromyxobacterdehalogenans
antiparasiticAntimalarial
Paenibacillin
Paenibacilluspolymyxa
anti-Gram+
Epidermin
Staphylococcusepidermidis Tü3298
anti-Gram+
Lichenicidin
Bacilluslicheniformis ATCC14580orVK21
anti-Gram+
EnterocinX
Enterococcusfaecium KU-B5
anti-Gram+
ThuricinCD
BacillusthuringiensisDPC6431
anti-Gram+
Duramycin
StreptoverticilliumgriseoverticillatumPA-48009
anti-Gram+
DuramycinB
StreptoverticilliumstrainR2075
anti-Gram+
DuramycinC
StreptomycesgriseoluteusR2107
anti-Gram+
Cinnamycin
StreptomycescinnamoneuscinnamoneusDSM40005
anti-Gram+antifungal
Mammaliancells
PAMP
PropionibacteriumjenseniiLMG3032
anti-Gram+
MichiganinA
Omatopathogen,Clavibactermichiganensissubsp.michiganensis
anti-Gram+
Ipomicin
sweetpotatopathogen, Streptomycesipomoeae
anti-Gram+
Actagardine
ActinoplanesgarbadinensisATCC31048and31049
anti-Gram+
Sublancin168
Bacillussubtilis168
anti-Gram+
BovicinHJ50
StreptococcusbovisHJ50
anti-Gram+
Thermophilin1277
StreptococcusthermophilusSBT1277
anti-Gram+
PlantaricinW
Lactobacillusplantarum
anti-Gram+
StaphylococcinC55
StaphylococcusaureusC55
anti-Gram+
Gallidermin
Staphylococcusgallinarum(F16/P57)Tü3928
anti-Gram+
anti-Gram-
Epilancin15X
Staphylococcusepidermidis 15X154
anti-Gram+
EpilancinK7
StaphylococcusepidermidisK7
anti-Gram+
Epicidin280
StaphylococcusepidermidisBN280
anti-Gram+
MicrobisporicinA1
Microbisporasp.,strain107891
anti-Gram+
SmbB
StreptococcusmutansGS5
anti-Gram+
Haloduracin
Bacillushalodurans C-125
anti-Gram+
Cypemycin
Streptomyce
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