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parts图标Abstract
This project aims to reduce the carbon emission and increase carbon economy during the fermentation of Clostridium tyrobutyricum (C. tyrobutyricum). Phosphoketolase (F/Xpk) gene sequences well fitted for C. tyrobutyricum L319 were obtained by strain screening and sequence optimization. Plasmids of the gene were constructed and transferred into C. tyrobutyricum L319 to construct non-oxidative glycolysis (NOG) pathway. Unlike the native Embden-Meyerhof-Parnas (EMP) pathway which emits CO2 during glycolysis, this alternative NOG pathway has no carbon emission. We constructed nine parts, BBa_K4886000- BBa_K4886008to express phosphoketolase using different F/Xpk sequences and promoters and construct NOG pathway in C. tyrobutyricum. The constructed NOG pathway was evaluated by experiments such as HPLC and fermentation experiments.


No.

Name

Type

Description

Length

1

BBa_K4886000

Basic

F/Xpk(QS), optimized F/Xpk gene derived from Bifidobacterium adolescentis ATCC 15703

2478

2

BBa_K4886001

Composite

Pthl-F/Xpk(BD), expression of F/Xpk from Clostridium acetobutylicum ATCC824 with Pthl promotor

3036

3

BBa_K4886002

Composite

Pthl-F/Xpk(QS), expression of optimized F/Xpk from Bifidobacterium adolescentis ATCC 15703 with Pthl promotor

3123

4

BBa_K4886003

basic

Pfba, derived from Clostridium tyrobutyricum

300

5

BBa_K4886004

composite

Pfba-F/Xpk(BD), expressing F/Xpk from Clostridium acetobutylicum with fba promotor

2739

6

BBa_K4886005

basic

Ptkt, promoter derived from Clostridium tyrobutyricum

 

300

7

BBa_K4886006

composite

Ptkt-F/Xpk(BD)

It is a part that is responsible for expressing F/Xpk from Clostridium tyrobutyricum. with tkt promotor.

2739

8

BBa_K488600 7

basic

F/ Xpk(ASR)

2433

 

9

BBa_K4886008

Composite

Pthl-F/Xpk (ASR)

3036