Heterologous overexpression of a novel delta-4 desaturase gene from the marine microalga Pavlova viridis in Escherichia coli as a Mistic fusion

Xu, Youzhi; Ning, Yan; Kong, Jing

doi:10.1007/s11274-011-0776-5

Cited by 7 publications

(7 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the case of several multidrug transporters and KDEL receptors, their expression in L. lactis led to growth ceasing almost instantly, resulting in little or no protein expression (Kunji et al, 2003). Although pkjDes4 was also shown to be seriously detrimental to host bacteria growth (Xu et al, 2011), the optical density (OD) of induced NZ9000/pNMDes4 culture was only slightly lower than that of NZ9000/pNM110 (Table S1). Therefore, it was deduced that successful pkjDes4 overexpression in L. lactis was at least partly due to the capacity of Mistic to circumvent growth impairment in lactococcal cells.…”

Section: Discussionmentioning

confidence: 99%

“…The eukaryotic membrane protein pkjDes4 was recently identified in Pavlova viridis (Xu et al, 2011). This protein and its family, namely microalgal front-end fatty acid desaturases, have not, to our knowledge, been overexpressed in L. lactis so far.…”

Section: Overexpression Of Pkjdes4 In L Lactismentioning

confidence: 97%

“…Fatty acid D4 desaturase is a family of eukaryotic transmembrane proteins responsible for the DHA synthesis in several species of marine microalgae (Tonon et al, 2003;Zhou et al, 2007), with a homologue (pkjDes4) recently identified in Pavlova viridis (Xu et al, 2011). For CLA production, a specific linoleate isomerase (LI) was first isolated and sequenced in Lactobacillus reuteri in 2004 by Rosson et al (2004).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Improved membrane protein expression in Lactococcus lactis by fusion to Mistic

Kong

2013

Microbiology

Self Cite

View full text Add to dashboard Cite

Difficulty overexpressing (eukaryotic) membrane proteins is generally considered as the major impediment in their structural and functional research. Lactococcus lactis possesses many properties ideal for membrane protein expression. In order to investigate membrane protein expression in L. lactis, we created a novel expression system by introducing Mistic, a short peptide previously identified in Bacillus subtilis, into L. lactis. The potential of this system was demonstrated in the overexpression of a eukaryotic membrane protein (pkjDes4) and a prokaryotic membrane protein (pkjLi), a newly isolated linoleate isomerase from Lactobacillus acidophilus. The expression levels reached up to 4.4 % and 45.2 % for pkjDes4 and pkjLi, respectively, which represented an exceptionally robust ability to overproduce membrane proteins. Moreover, the expressed pkjLi was functional, with its catalysing nature characterized for the first time in this species. Up to 0.852 mg ml "1 conjugated linoleic acid was obtained during the linoleic acid conversion catalysed by the recombinant lactococcal strains. In summary, we established a membrane protein expression system in L. lactis and examined its functionality. Our results demonstrate that the Mistic chaperoning strategy can be efficiently applied to L. lactis hosts and show its extraordinary capacity to facilitate the high-yield production of intractable membrane proteins.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Overexpression Of Pkjdes4 In L Lactismentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Improved membrane protein expression in Lactococcus lactis by fusion to Mistic

Kong

2013

Microbiology

Self Cite

View full text Add to dashboard Cite

show abstract

“…Feeding trials with non-native substrates [48] Pavlova salina PsD4 Eukaryotic DES expressed in E. coli Utilization of the novel protein fusion MISTIC [ 160] Streptomyces platensis…”

Section: Post-translational Regulation Of Desmentioning

confidence: 99%

Integral Membrane Fatty Acid Desaturases: A Review of Biochemical, Structural, and Biotechnological Advances

Nachtschatt

Okada

Speight

2020

Euro J Lipid Sci & Tech

View full text Add to dashboard Cite

Sustainably produced platform chemicals are increasingly sought after. Fatty acids containing olefinic and hydroxy groups are particularly interesting as platform chemicals for regio-and stereo-selective downstream reactions. Integral membrane fatty acid desaturases can selectively introduce double bonds or catalyze unusual reactions such as acetylenation, conjugation, hydroxylation, and epoxidation. Here, a systematic overview of recent achievements and key literature of integral membrane desaturases is provided in the context of biotechnological potential of the enzyme family. Practical Application: Integral membrane fatty acid desaturases may be the most promising tool to increase the scope of fatty acids as platform chemicals. Plant oil production is well researched and an established sustainable pathway toward high-yield chemicals. In addition, effort is increasing toward creating microbial high-yield oil producers. The current bottleneck is the limited fatty acid species that can be produced at high yields. It is envisioned that future enzyme engineering work will enable the targeted design of desaturases to produce modified fatty acids that follow industry needs. This review provides a systematic overview of the natural diversity and complexity of integral membrane fatty acid desaturases to help researchers expand existing fatty acid production beyond currently utilized fatty acid species.

show abstract

“…It was isolated from Thraustochytrium sp. (Qui et al 2001;Zank et al 2005), Schizochytrium aggregatum (Kinney et al 2004), Isochrysis galbana (Pereira et al 2004), Thalassiosira pseudonana (Tonon et al 2005), Pavlova viridis (Xu et al 2011). It has typical sign for 'front-end' desaturase -Q in 3 rd His box instead of H, cytochrome b 5 domain on N-terminal, but in 2 nd His box contains one more amino acid residue (Fig.…”

Section: δ4-desaturasementioning

confidence: 99%

Characterization of membrane-bound fatty acid desaturases

Klempová

Mihálik²,

Čertı́k³

2013

gpb

View full text Add to dashboard Cite

Abstract. Membrane-bound desaturases play key role in metabolism of polyunsaturated fatty acids. Characterization of these enzymes and their genes is the first step in basic understanding of their proper functioning in living cells as well as in tailor-made preparation of highly-specific and highlyproductive strains of microorganisms interesting for applied biotechnology. It is also the crucial step in creation of transgenic agricultural crops with enhanced content of individual polyunsaturated fatty acids. Properties and applications of identified membrane-bound desaturases genes and enzymes are discussed in this review.

show abstract

Heterologous overexpression of a novel delta-4 desaturase gene from the marine microalga Pavlova viridis in Escherichia coli as a Mistic fusion

Cited by 7 publications

References 19 publications

Improved membrane protein expression in Lactococcus lactis by fusion to Mistic

Improved membrane protein expression in Lactococcus lactis by fusion to Mistic

Integral Membrane Fatty Acid Desaturases: A Review of Biochemical, Structural, and Biotechnological Advances

Characterization of membrane-bound fatty acid desaturases

Contact Info

Product

Resources

About