CSU Bakersfield Researcher Earns Accolades

Alexander Candia went from satisfying people's hunger to satisfying his own thirst for knowledge, and along the way became one of the top researchers at California State University, Bakersfield.

Candia is a senior chemistry major who plans to continue his studies to eventually become a physician's assistant. His work with chemistry professor Roy LaFever, focusing on natural substances of the horehound plant, earned him first place awards in both the CSUB Student Research Competition and also the regional research competition at CSU Fresno.

Horehound, a member of the mint family, has been used for centuries to relieve respiratory and bronchial ailments, and a compound known as marrubiin has been implicated as the active constituent. Marrubiin is a member of a family of natural products known as diterpenes, and shares similarities with the anti-cancer diterpene taxol. Earlier research indicates marrubiin has some interesting biological effects including anti-viral and antibiotic activities. Extracts from the plant also repel a number of insect pests making it a promising lead for use as an insecticide.

LaFever set out to find how and where marrubiin is produced in the plant, and last year found what he was looking for.

"There are specialized structures on mint leaves that produce the aromatic oils," LaFever said. "So we looked at the horehound plant to see if it had these specific leaf structures, and found that it did."

The leaves have a fuzzy texture to them, he knew, and found what he was looking for below the forest of hairs on the leaf. "Underneath the fur are clusters of cells that include a cylindrical layer where the compound is made," he said. These structures are only visible with the aid of a microscope and are roughly 100-times smaller than a flea. "We developed a technique to isolate the specialized structures and analyze the contents directly. Using the technique it was found that the marrubiin was indeed produced and stored in these structures known as secretory glands.

"We not only know that marrubiin is made there, but we can also isolate the proteins and enzymes from the cells that are responsible for making marrubiin.

"The ability to isolate these glands in relatively pure form is an exciting development."

Marrubiin is related to the fragrant aromas in mint known as terpenoids, but is chemically distinct, he said. Marrubiin is also related to taxol, found in the Western Yew tree, which is "used to treat ovarian cancer.

When we find the gene that produces marrubiin, we'll be able to produce it in large quantity and begin to modify its structure in hopes of developing taxol-like anti-cancer activity and improved antibiotics."

That's where Candia came in. Since LaFever and previous student researchers had discovered where the marrubiin was produced, Candia's task was to figure out how to produce more of it.

"What I did was manipulate two metal ions - magnesium and manganese - that are responsible for the plant's metabolism," Candia said. His experiment involved feeding one collection of plants Miracle Grow fertilizer, and another the fertilizer supplemented with magnesium and manganese. The sample treated with both metals had enhanced growth and a significant increase in marrubiin production. We were able to increase the marrubiin production in treated plants by a factor of 30. That is the treated plants produced 30 time more marrubiin than those without the additional magnesium and manganese"

And while Candia is now able to mass-produce marrubiin, at one point he was responsible for mass-producing meals for banquets. "I went to chef school originally," the 36-year-old said. "I was a chef and worked at Bakersfield Country Club."

But by the mid-1990s, he yearned to do something else, and in 1996 enrolled at CSUB, taking one class per quarter. "My wife finally said let's bite the bullet, go full time. Now I work weekends and go to school
during the week.

"I actually started in pre-med. I loved being in the lab. It was like being in the kitchen. Since I was involved in the Chemistry Club I was working on a project. Dr. LaFever told me I had aptitude in the lab and suggested I apply for a scholarship. I did, and got the scholarship, and now I get to work in the lab all the time. I love to work in the lab, and can't believe how fast the time goes."

LaFever is delighted with Candia's success. Not only does it help advance the research LaFever has been pursuing for several years, but also it helps Candia in his pursuit of a career.

"Alex has really done a first-rate job," LaFever said. "He is a very determined student who has developed a keen interest in biochemical research, and possesses a variety of laboratory skills normally learned in graduate studies. I am very pleased that Alex has been awarded a Student Research Scholarship for the 2001-02 academic year. His upcoming research will focus on manipulating metals supplied to yew trees to evaluate taxol production under different conditions. This is an exciting project that fits well with the on-going effort and extends our previous findings."