Feeding strategy design for recombinant human growth hormone production by Bacillus subtilis

Abstract

Defined and semi-defined medium-based feeding strategies were developed to enhance recombinant human growth hormone (rhGH) production by Bacillus subtilis BGSC-1A178 (scoC (-)) strain carrying pMK4::pre(subC)::hGH. Defined medium-based feeding strategies were designed by exponential feeding of glucose and (NH4)2HPO4 at two pre-determined specific growth rates, µ 0 = 0.10 and 0.17 h(-1). Semi-defined medium-based feeding strategies were designed by exponential feeding of substrate solution consisting of glucose, (NH4)2HPO4, peptone, and trace salt solution (PTM1) at three pre-determined specific growth rates, µ 0 = 0.10, 0.17, and 0.25 h(-1). At all the strategies applied, transition cultivation time from batch to fed-batch operation was t T = 4 h. The highest rhGH concentration was obtained as C rhGH = 0.5 g L(-1) with semi-defined medium-based feeding strategy designed with µ 0 = 0.25 h(-1) using feed substrate stock solution containing 200 g L(-1) glucose, 117 g L(-1) (NH4)2HPO4, 100 g L(-1) peptone, and 5 mL L(-1) PTM1 at t = 22 h when the cell concentration reached to C X = 8.29 g L(-1). The overall product and cell yields on glucose were obtained as [Formula: see text] = 7.21 mg g(-1) and [Formula: see text] = 0.12 g g(-1), respectively. The results indicate the requirement of designing continuous feed stream in fed-batch production to enhance rhGH production by r-B. subtilis.

Bacillus subtilis-Based Probiotic Improves Skeletal Health and Immunity in Broiler Chickens Exposed to Heat Stress

“…Bacillus subtilis has been documented to have broadly biochemical effects such as antimicrobial, anti-inflammatory, antioxidant, enzymatic, and immunomodulatory activities in humans and various animals [ 52 , 177 ]. Bacillus subtilis also releases several neuropeptides, such as human-like growth hormone [ 178 , 179 ], parathyroid hormone [ 180 ], and tryptophan [ 181 , 182 , 183 ]. All these biochemical compounds regulate bone development, especially tryptophan, the precursor of serotonin biosynthesis, which acts on the central serotonergic system due to the fact that it can pass the blood–brain barrier [ 184 ].…”

Let There Be Light: Genome Reduction Enables Bacillus subtilis to Produce Disulfide-Bonded Gaussia Luciferase

“…The efficiency of protein secretion in Bacillus depends on diverse parameters, including the properties of the POI and the signal peptide (SP) used to direct it into the Sec pathway, , the type of promoter used for expression, − the cultivation conditions, the modulation of particular chaperones and secretion pathway components, − or combinations thereof. , Although machine learning approaches to select optimal SPs in silico are currently being developed, , the identification of suitable SPs still requires experimental testing in the laboratory.…”

Design of a high‐efficiency synthetic system for l‐asparaginase production in Bacillus subtilis

“…Promoters greatly influence the transcription levels of genes. A large number of promoters have previously been used successfully to improve product yield . By replacing the promoter, the production increased from 12% to five times as much .…”

“…Promoters greatly influence the transcription levels of genes. A large number of promoters have previously been used successfully to improve product yield [14][15][16][17][18].…”