High tetramethylpyrazine production by the endophytic bacterial Bacillus subtilis isolated from the traditional medicinal plant Ligusticum chuanxiong Hort.

Abstract

Tetramethylpyrazine (TMP) with significant protective effects on cardiovascular is the active ingredient of traditional Chinese medicine Rhizoma Chuanxiong (RC). However, many studies have reported the low content of TMP in RC. The endophytes of medicinal plants have the biosynthetic potential to produce the same or similar active metabolites as the host, while few reports were conducted to explore the endophytic bacteria of Ligusticum chuanxiong Hort. and its productive capacity for the important ingredient TMP. The present paper focuses on the isolation and identification of TMP producing endophytic bacteria from RC. In this study, the endophytic bacteria were isolated from the rhizome of Ligusticum chuanxiong Hort. (Umbelliferae). Yeast extract peptone glucose medium (YP) was used for fermentation medium (37 °C, 220 rpm agitation, 144 h). GC and GC/MS were performed to determine and verify the product, the fermentation characteristics were investigated. Morphological observation, physiological and biochemical indexes combining with 16S rRNA sequence analysis were carried out to identify the endophytic bacteria. As a result, five strains of endophytic Bacillus subtilis were firstly isolated and identified from RC, named as LB3, LB3-2-1, LB6-2, LB4, LB5 respectively. All five strains of endophytic B. subtilis produced TMP, while LB5 had the highest production of 10.69 g/L at the 144 h fermentation. This work demonstrates the fact that the endophytic B. subtilis of RC can produce a high level of TMP, indicating the endophytic B. subtilis might play a role in the accumulation of TMP during the growth period of RC.

Production of tetra-methylpyrazine using engineered Corynebacterium glutamicum

“…This report illustrates two parameters by which C. glutamicum can be modified to produce tetra-methylpyrazine, TMP. Our highest final titers from batch and fed-batch production modes compare favorably with published titers from Bacillus subtilis (Xiao et al., 2014; Yin et al., 2018) and do not require a post-cultivation denaturing step to produce TMP . Unlike E .…”

“Breeding on Mountains” Resulted in the Reorganization of Endophytic Fungi in Asexually Propagated Plants (Ligusticum chuanxiong Hort.)

“…It has been found that the endophytic fungi of L. chuanxiong are often significantly related to their active ingredients. For example, the content of ibutilide is significantly related to endophytes, such as Penicillium and Wardell , and Bacillus subtilis can secrete Ligustrazine ( Yin et al, 2018a , b , 2019 ). Therefore, endophytic fungi can affect the quality of the rhizome of L. chuanxiong by regulating the production of its secondary metabolites.…”

Analysis of Chuanxiong Rhizoma substrate on production of ligustrazine in endophytic Bacillus subtilis by ultra high performance liquid chromatography with quadrupole time‐of‐flight mass spectrometry

“…According to the previous preliminary study of our research, a considerable yield of ligustrazine (10.69 g/L, 144 h) can be produced by the endophytic B.subtilis LB5, which was isolated from the fresh rhizome of Ligusticum chuanxiong Hort [24]. The yeast extract peptone glucose medium provided an excellent and nutritious growing environment for the fermentation of B. subtilis LB5.…”