Team:SZPT-CHINA/Parts/Overview


part_overview
PARTS
INTRODUCTION
Our parts consist of three parts: (1) related to acid sensors; (2) related to the expression of antihypertensive peptides; (3) autolysis of lactic acid bacteria. All parts were generated during our project. There are 12 basic parts and 4 composite parts.
PARTS RELATED TO PH SENSOR
The rcfB promoter, a member of the CRP-FNR positive regulation family, was amplified from chromosomal DNA of L. Lactis subsp. lactis IL1403. It is an important part of the P170 promoter regulated by acid. When the external PH value declines to 5.5, rcfb promoter will be highly upregulated. [1]
Lac repressor (LacR) and operator (O1 and O2) sequences is part of the lactose operon derived from Lactobacillus delbrueckii. O1 and O2 may serve as operators for the binding of the lac repressor (LacR). [2]
Part Number
Name
Type
Part Description
BBa_K3142000 PrcfB Basic Acid-induced promoter
BBa_K3142001 O1/O2 Basic Lactose operator
BBa_K3142015 LacR Basic Lac repressor
BBa_K3142002 PrcfB+LacR Composite Acid-induced promoter+ repressor LacR of lac operon
PARTS RELATED TO EXPRESSION OF ANTIHYPERTENSIVE PEPTIDE
Antihypertensive peptides (AHPs) are protein-derived short peptides released by hydrolysis processes, which act as inhibitors of angiotensin-converting enzyme and renin.
The 11S globulin from amaranth (A11Sg) is the most abundant storage protein in mature seeds and is well recognized for its nutritional value. We used this globulin to engineer a new protein by adding five highly active antihypertensive peptides at its C-terminal end to improve its functionality. [3]
Part Number
Name
Type
Part Description
BBa_K3142003 FR Basic Arginine Linker
BBa_K3142004 A11Sg Basic 11S globulin from amaranth
BBa_K3142005 VY Basic Antihypertensive peptide
BBa_K3142006 IPP Basic Antihypertensive peptide
BBa_K3142007 VPP Basic Antihypertensive peptide
BBa_K3142008 KYLCY Basic Antihypertensive peptide
BBa_K3142009 FKGKYYP Basic Antihypertensive peptide
BBa_K3142010 AHPM Composite Antihypertensive peptide multimer
BBa_K3142011 A11Sg-AHPM Composite Complex of 11S globulin from amaranth and Antihypertensive peptide multimer
PARTS RELATED TO LACTIC ACID BACTERIA AUTOLYSIS
Enhanced truncated promoters (T-αcrp) which include a modified enhancer element from the CC(-41.5)α(-63) synthetic elements upstream of the T-crp promoters. Minimal “truncated” promoters (T-crp) containing ~ 50 bp upstream of the transcription start site with care taken to include the full CRP binding site. T-αcrp promoter can response to glucose starvation to Initiate expression of downstream genes. [4]
N-acetylmuramidase (acmA) from Lactobacillus acidophilus NCFM dissolves the cell wall of lactic acid bacteria, causing lactic acid bacteria to cleave. [5]
Part Number
Name
Type
Part Description
BBa_K3142012 PT-αCRP Basic Enhanced truncated promoters
BBa_K3142013 acmA Basic N-acetylmuramidase
BBa_K3142014 PT-αCRP + acmA Composite Enhanced truncated promoters+ N-acetylmuramidase
References
[1] Ismail AKYOL, Ugur COMLEKCIOGLU, Asuman KARAKAS,et,al. Regulation of the acid inducible rcfB promoter in Lactococcus lactis subsp. Lactis. Annals of Microbiology, 58 (2) 269-273 (2008)
[2] United States Patent . Patent No.US 6,929,931 B1. Date of Patent:Aug. 16, 2005
[3] Edgar Espinosa-Hernández , Jocksan Ismael Morales-Camacho , D. Alejandro Fernández-Velasco,et,al. The insertion of bioactive peptides at the C-terminal end of an 11S globulin changes the structural stability and improves the antihypertensive activity. Electronic Journal of Biotechnology 37 (2019) 18–24
[4] William Henry Bothfeld, Grace Kapov, and Keith Tyo. A glucose-sensing toggle switch for autonomous, high productivity genetic control. ACS Synth. Biol,Just Accepted Manuscript•DOI: 10.1021/acssynbio.6b00257•Publication Date (Web): 08 Mar 2017
[5] Pfeiler EA, Azcarate-Peril MA, Klaenhammer TR. Characterization of a novel bile-inducible operon encoding a two-component regulatory system in Lactobacillus acidophilus. J Bacteriol. 2007 Jul;189(13):4624-34. Epub 2007 Apr 20.