Autoinduction, purification, and characterization of soluble α-globin chains of crocodile (Crocodylus siamensis) hemoglobin in Escherichia coli

Kabbua, Thai; Anwised, Preeyanan; Boonmee, Atcha; Subedi, Bishnu P.; Pierce, Brad S.; Thammasirirak, Sompong

doi:10.1016/j.pep.2014.08.013

Cited by 9 publications

(7 citation statements)

References 41 publications

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“…Based on the quantitative result, the incorporated heme in purified porcine MG could reach 0.22 mol of heme/mol of MG (Figure 4D), which is significantly higher than the content of heme (0.12 mol of heme/mol α-globin) expressed by E. coli. 42 In summary, the porcine MG can be efficiently secreted in recombinant K. phaffii X33-α GAP-MG strain by α-factor secretory signal peptide and G1 promoter. The highest titer and productivity of porcine MG reached 285.42 mg/L and 2.90 mg/(L h), respectively, in fed-batch fermentations using modified BMGY medium with the supplement of 150 mg/L of hemin at 30% DO-stat and 30 °C.…”

Section: ■ Results and Discussionmentioning

confidence: 94%

“…Dependent on the purified porcine MG, the content of heme was examined because incorporated heme in MG is closely related to the color and flavor in meat products and is the main source of iron in the human diet . At first, the pyridine hemochromagen assay used to determine the content of heme in hemoglobin was applied to examine the heme in porcine MG . However, there was no obvious peak at 500–600 nm for the purified MG.…”

Section: Resultsmentioning

confidence: 99%

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Efficient Secretory Expression and Purification of Food-Grade Porcine Myoglobin in Komagataella phaffii

Zhang

Zhao

Wang

et al. 2021

J. Agric. Food Chem.

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Myoglobin (MG) is one of the eukaryotic heme-binding proteins that is closely associated with the real color and metallic taste of meat and can be used as a color additive in artificial meat alternatives. However, the traditional extraction methods are expensive and time-consuming and the heterologous biosynthesis of MG has never been reported. Herein, we achieved the secretory expression of porcine MG by engineered Komagataella phaffii using the suitable host (X33), signal peptide (α-factor signal peptide), and modified constitutive promoter (G1 promoter). In addition, the fermentation conditions for MG production were optimized at shaking-flask level (BMGY medium with 40 mg/L of hemin, 30 °C) and at fermenter level (30% DO, feeding 150 mg/L of hemin), resulting in the highest titer of 285.42 mg/L MG in fed-batch fermentations. Furthermore, a purification method for food-grade MG was developed, which can obtain 0.22 mol of heme/mol of MG with 88.0% purity and 66.1% recovery rate.

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Section: ■ Results and Discussionmentioning

confidence: 94%

Section: Resultsmentioning

confidence: 99%

Efficient Secretory Expression and Purification of Food-Grade Porcine Myoglobin in Komagataella phaffii

Zhang

Zhao

Wang

et al. 2021

J. Agric. Food Chem.

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“…Kabbua et al. found that recombinant α‐globin from cHb can act as a heme‐nitric oxide and/or oxygen‐binding (H‐NOX) hemoprotein. Furthermore, the value of NO inhibition activity of CHH was higher than yellowfin tuna hydrolysate .…”

Section: Resultsmentioning

confidence: 99%

Novel antioxidant and anti‐inflammatory peptides from the Siamese crocodile (Crocodylus siamensis) hemoglobin hydrolysate

Lueangsakulthai

Phosri

Theansungnoen

et al. 2017

Biotech and App Biochem

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Novel antioxidant and anti-inflammatory peptides were isolated from hydrolysates of Siamese crocodile (Crocodylus siamensis) hemoglobin. C. siamensis hemoglobin hydrolysates (CHHs) were obtained by pepsin digestion at different incubation times (2, 4, 6, and 8 H) at 37 °C and subjected to antioxidant and anti-inflammatory activity assessment. CHH obtained by 2-H hydrolysis (2H-CHH) showed the highest anti-inflammatory activity with respect to decreasing nitric oxide (NO) production, whereas the strongest antioxidant activity was found for 6-H hydrolysis (6H-CHH) against nitric oxide radicals. To evaluate the anti-inflammatory and antioxidant activity of individual peptide components, 2H-CHH and 6H-CHH were purified by semipreparative HPLC. Peptide fraction P57 isolated from 6H-CHH was found to exhibit the highest nitric oxide radical inhibition activity (32.0%). Moreover, purification of 2H-CHH yielded peptide fraction P16, which displayed a high efficacy in decreasing NO production of macrophage RAW 264.7 cells (83.2%) and significantly reduced proinflammatory cytokines and inflammatory mediators interleukin-6 (IL-6), interleukin-1 beta (IL-1β), and prostaglandin-E2 (PGE ) production to about 2.0, 0.3, and 1.9 ng/mL, respectively. Using LTQ orbitrap XL mass spectrometry, active peptide sequences were identified as antioxidant KIYFPHF (KF7), anti-inflammatory SAFNPHEKQ (SQ9), and IIHNEKVQAHGKKVL (IL15). Additionally, CHHs simulated gastric and intestinal in vitro digestion positively contributed to antioxidant and anti-inflammatory activity. Taken collectively, the results of this work demonstrate that CHHs contain several peptides with anti-inflammatory and antioxidant properties, which may prove valuable as treatment or supplement against diseases associated with inflammation and oxidative stress.

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“…Usually haemoglobin can bind (carry) up to four oxygen molecules via a haem group consists of an iron (Fe) ion (charged atom) held in a heterocyclic ring, known as a porphyrin (Ali et al 2013). Because of the ability of haemoglobin that can carry not only oxygen but also other molecules such as in recent studied found that recombinant a-globin from C. siamensis haemoglobin can act as a haem-nitric oxide and/or oxygen binding (H-NOX) haemoprotein (Kabbua et al 2014). Therefore, the antibacterial mechanism was investigated with focus on iron homeostasis and oxidative stress, which in numerous pathogens is associated with host haem-iron availability.…”

Section: Discussionmentioning

confidence: 99%

A novel antibacterial peptide derived fromCrocodylus siamensishaemoglobin hydrolysate induces membrane permeabilization causing iron dysregulation, oxidative stress and bacterial death

et al. 2017

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Autoinduction, purification, and characterization of soluble α-globin chains of crocodile (Crocodylus siamensis) hemoglobin in Escherichia coli

Cited by 9 publications

References 41 publications

Efficient Secretory Expression and Purification of Food-Grade Porcine Myoglobin in Komagataella phaffii

Efficient Secretory Expression and Purification of Food-Grade Porcine Myoglobin in Komagataella phaffii

Novel antioxidant and anti‐inflammatory peptides from the Siamese crocodile (Crocodylus siamensis) hemoglobin hydrolysate

A novel antibacterial peptide derived fromCrocodylus siamensishaemoglobin hydrolysate induces membrane permeabilization causing iron dysregulation, oxidative stress and bacterial death

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