Steamed bread is a traditional staple food in China, and it has gradually become loved by people all over the world because of its healthy production methods. With the improvement in people’s living standards, the light flavor of steamed bread fermented by single yeast cannot meet people’s needs. Multi-strain co-fermentation is a feasible way to improve the flavor of steamed bread. Here, the dynamic change profiles of volatile substances in steamed bread co-fermented by Saccharomyces cerevisiae SQJ20 and Wickerhamomyces anomalus GZJ2 were analyzed using the electronic nose (E-nose) and headspace solid-phase microextraction combined with gas chromatography–mass spectrometry (HS-SPME-GC-MS). The five detectors of the E-nose rapidly detected the changes in volatile substances in different dough or steamed bread with the highest response value in co-fermented dough. A total of 236 volatile substances were detected in all the samples using HS-SPME-GC-MS, and alcohols were the most variable component, especially Phenylethyl alcohol. Significantly, more alcohols and esters were upregulated in co-fermented dough, and the addition of W. anomalus GZJ2 improved the key volatile aroma compounds of steamed bread using the relative odor activity value method (ROAV), especially the aldehydes and alcohols. Moreover, these key volatile aroma compounds can be quickly distinguished using the W2S detector of the E-nose, which can be used for the rapid detection of aroma components in steamed bread.

In this study, an efficient mutagenesis and rapid screening method of high-yield gellan gum mutant by atmospheric and room temperature plasma (ARTP) treatment combined with Near-Infrared Spectroscopy (NIRS) was proposed. A NIRS model for the on-line detection of gellan gum yield was constructed by joint interval partial least squares (siPLS) regression on the basis of chemical determination and NIRS acquisition of gellan gum yield. Five genetically stable mutant strains were screened using the on-line NIRS detection of gellan gum yield in the fermentation from approximately 600 mutant strains induced by ARTP. Remarkably, compared with the original strain, the gellan gum yield of mutant strain 519 was 9.427 g/L (increased by 133.5%) under the optimal fermentation conditions, which was determined by single-factor and response surface optimization. Therefore, the method of ARTP mutation combined with the NIRS model can be used to screen high-yield mutant strains of gellan gum and other high-yield polysaccharide strains.

Pseudomonas plecoglossicida JUIM01 is an industrial 2-keto-d-gluconate (2KGA)-producing strain. However, its regulation mechanism of 2KGA metabolism remains to be clarified. Among other reported Pseudomonas species, the 2-ketogluconate utilization operon (kgu operon) plays key roles in 2KGA catabolism. In this study, the structural genes of the kgu operon and its promoter in P. plecoglossicida JUIM01 were identified using reverse transcription PCR and lacZ reporter gene fusion. The results showed the kgu operon in P. plecoglossicida was composed of four structural genes: kguE, kguK, kguT, and kguD. The ptxS gene located upstream of kguE was excluded from the kgu operon. Then, the knockout and corresponding complementation strains of kguE, kguK, kguT, and kguD were constructed, respectively, to prove the kgu operon was involved in 2KGA catabolism of P. plecoglossicida. The knockout stains, especially JUIM01ΔkguE, showed potential as industrial production strains for 2KGA. Moreover, the transcriptional regulation mechanism of PtxS on the kgu operon was elucidated using multiple methods. In P. plecoglossicida, the LacI-family transcription regulator PtxS could recognize a 14 bp palindrome (5′-TGAAACCGGTTTCA-3′) within the promoter region of the kgu operon and specifically bind to a 26 bp region where the palindrome was located. As the binding sites overlapped with the transcription start site of the kgu operon, the binding of PtxS possibly hindered the binding of RNA polymerase, thus repressing the transcription of the kgu operon and further regulating 2KGA catabolism. 2KGA bound to PtxS as an effector to dissociate it from the kgu operon promoter region, so as to relieve the transcription repression. The results will provide strategies for improving the product accumulation in 2KGA industrial production and theoretical bases for the construction of a Pseudomonas chassis.